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The DNA Damage Response Induces IFN
Sabrina Brzostek-Racine, Chris Gordon, Sarah Van Scoy and Nancy C. Reich
J Immunol November 15, 2011, 187 (10) 5336-5345; DOI: https://doi.org/10.4049/jimmunol.1100040
Sabrina Brzostek-Racine
Department of Molecular Genetics and Microbiology, Stony Brook University, Stony Brook, NY 11794
Sarah Van Scoy
Nancy C. Reich
This study reveals a new complexity in the cellular response to DNA damage: activation of IFN signaling. The DNA damage response involves the rapid recruitment of repair enzymes and the activation of signal transducers that regulate cell-cycle checkpoints and cell survival. To understand the link between DNA damage and the innate cellular defense that occurs in response to many viral infections, we evaluated the effects of agents such as etoposide that promote dsDNA breaks. Treatment of human cells with etoposide led to the induction of IFN-stimulated genes and the IFN-α and IFN-λ genes. NF-κB, known to be activated in response to DNA damage, was shown to be a key regulator of this IFN gene induction. Expression of an NF-κB subunit, p65/RelA, was sufficient for induction of the human IFN-λ1 gene. In addition, NF-κB was required for the induction of IFN regulatory factor-1 and -7 that are able to stimulate expression of the IFN-α and IFN-λ genes. Cells that lack the NF-κB essential modulator lack the ability to induce the IFN genes following DNA damage. Breaks in DNA are generated during normal physiological processes of replication, transcription, and recombination, as well as by external genotoxic agents or infectious agents. The significant finding of IFN production as a stress response to DNA damage provides a new perspective on the role of IFN signaling.
An effective DNA damage response is critical for maintaining genomic integrity and preventing mutations that can lead to cancer. Double-strand breaks are the most severe lesions, and they can occur during DNA replication, lymphocyte V(D)J gene rearrangement, meiosis, viral infection, and in response to naturally occurring ionizing radiation (1–5). These DNA breaks are sensed rapidly, and accurate repair is essential to prevent permanent genomic damage. However, the cellular response to DNA damage engages more than just DNA repair machinery; it engages complex signaling pathways that can promote cell survival or cell death (6, 7). In this report, the activation of an additional network is revealed: the IFN signal pathway.
A primary transducer of the response to double-strand breaks is the nuclear kinase ataxia-telangiectasia mutated (ATM) (8). ATM belongs to a family of PI3K-related kinases, several of which are involved in the DNA damage response, including ATM and Rad3 related (ATR) and DNA-dependent protein kinase (DNA-PK) (9). ATM transduces the DNA damage response signal by phosphorylating downstream effectors such as the checkpoint kinases Chk1 and Chk2 and the p53 tumor suppressor. These effectors in turn establish cell-cycle arrest to allow repair of damaged DNA or promote damage-induced apoptosis. Major alterations in gene expression occur during this time, and this reflects the action of not only p53, but also other transcription factors (10). One transcription factor that is activated in response to DNA damage is NF-κB (11). NF-κB regulates the expression of diverse genes involved in cellular responses that include survival, proliferation, tissue remodeling, inflammation, immunity, and stress. We have found that NF-κB activation in response to DNA damage directs the induction of the IFN system, a stress pathway best known for its ability to confer viral resistance.
IFNs play vital roles in both innate and adaptive immunity and consist of three families of cytokines that bind to distinct cell-surface receptors and are designated types I, II, and III (12). The genes encoding type I IFN (primarily α and β) and type III (λ) IFN are induced in response to viral or bacterial infection (13). The single type II (γ) IFN gene is induced primarily following receptor activation of T cells and NK cells. The regulated expression of the IFN-β gene in response to viral infection is a paradigm for cooperativity of DNA binding factors (14). NF-κB and IFN regulatory factors (IRFs) function along with activating transcription factor-2/c-Jun in the IFN-β enhancer. The IRFs were first characterized as regulators of type I IFN genes and IFN-stimulated genes (ISGs) and are now known to have diverse roles in immunity (15). Activation of ubiquitous IRF-3 during viral infection supports induction of a subset of ISGs and the IFN-β and IFN-α genes (16, 17). The IRF-1 and IRF-7 genes are induced in response to secreted IFN and can play a role in the secondary response to IFN (18). IFNs bind to cell-surface receptors that activate Janus kinases and the tyrosine phosphorylation of STAT1 and STAT2 (19, 20). We report in this study that signaling via the DNA damage response in human cells primarily induces the IFN-λ and IFN-α genes. The promoters of the IFN-λ genes have been found to possess both IRF and NF-κB binding sites (21, 22). We demonstrate that NF-κB activation in response to DNA damage is sufficient and necessary to induce human IFN-λ.
This study identifies IFN signaling as part of the DNA damage response. IFNs are essential components of innate immunity and are well recognized for their ability to inhibit viral infection and activate immune effector cells (23). In addition, they are known for their antitumor effects by inhibiting proliferation of cancerous cells and promoting apoptosis (24–26). The IFN arm of the DNA damage response may have evolved as an antiviral mechanism in reaction to DNA damage induced by viruses, as a mechanism that reduces cellular proliferation to allow DNA repair, or as a mechanism to promote the death of cells with irreparable damage.
Cell culture, transfections, and infections
HeLa S3, HT1080, and THP-1 cells were obtained from American Type Culture Collection. THP-1 cells were maintained in RPMI 1640 with 8% FBS; other cells were maintained in DMEM with 8% FBS. Primary human monocytes were isolated from blood cells of healthy donors (Long Island Blood Services) using the Monocyte Isolation Kit II (Miltenyi Biotec, Auburn, CA) and maintained in RPMI 1640 with 10% FBS. Stable HT1080 transfectants with tetracycline-inducible expression of IRF-7 were generated according to the manufacturer's instructions (T-Rex system; Invitrogen, Carlsbad, CA), and the gene was induced with 2 μg/ml doxycycline. Murine embryo fibroblasts (MEFs) from NF-κB essential modulator (NEMO)/IκB kinase (IKK) γ knockout mice were a gift of Dr. Kenneth Marcu (Stony Brook University) (27), MEFs from IRF3 knockout mice were a gift of Dr. Tadatsugu Tanaguchi (University of Tokyo) (16, 17), MEFs from IRF1 knockout mice were a gift of Dr. Tak Mak (University of Toronto) (28), and MEFs from IRF7 knockout mice were a gift of Dr. Michael Gale (University of Washington) (29). DNA transfections were performed using FuGENE 6 (Roche Diagnostics, Indianapolis, IN) or TransIt (Mirus, Madison, WI). Newcastle disease virus (NDV) (NJ-LaSota-1946) was a gift from Dr. Paula Pitha-Rowe (Johns Hopkins University, Baltimore, MD) and was propagated as described previously (30). Infections were performed at 200 hemagglutination units/ml.
Plasmids and luciferase assays
IRF-3, STAT1, and STAT2 constructs have been described (31–33). The dominant-negative IκBα plasmid (S32A/S36A) was a gift of Dr. Dean Ballard (Vanderbilt University, Nashville, TN) (34). The hemagglutinin (HA)-tagged ubiquitin–K63-only (HA-Ub0R63K) plasmid was a gift of Dr. Dafna BarSagi (New York University) (35). The reporter plasmid encoding the IFN-λ1 promoter regulating expression of the firefly luciferase gene (pλ1[−554/+14]Luc) was a gift of Dr. Takashi Fujita (Kyoto University, Kyoto, Japan) (21). The human IRF-7A gene was obtained from Dr. Joseph S. Pagano (University of North Carolina, Chapel Hill, NC) (36) and subcloned into the pcDNA4/TO/Myc-polyhistidine (His) vector (T-Rex system; Invitrogen). The Dual-Luciferase reporter assay system was used for luciferase assays with the Renilla luciferase construct pRL-null as an internal control (Promega, Madison, WI).
Etoposide was used at 40 μg/ml, and camptothecin, adriamycin, and mitomycin were used at the concentrations indicated (Sigma-Aldrich, St. Louis, MO). NF-κB inhibitor BAY 11-7085 was obtained from Alexis Biochemicals (San Diego, CA) and used at 5 μM, and the IKKβ inhibitor ML120B was a gift from Millennium Pharmaceuticals (Cambridge, MA) and used at 20 μM. The ATM inhibitor AZ12622702/KU55933 was a gift from AstraZeneca (Cheshire, U.K.) and used at 10 μM. Abs recognizing STAT2 (C-20), c-Myc epitope (sc-40), CREB-binding protein (CBP; A-22), and normal rabbit IgG were purchased from Santa Cruz Biotechnology (Santa Cruz, CA). Ab to STAT2 phosphorylated on tyrosine 689 was obtained from Upstate Biotechnology (Lake Placid, NY). Abs to p65 phosphorylated on serine 536 and STAT1 phosphorylated on tyrosine 701 were obtained from Cell Signaling Technology (Beverly, MA). Ab against HA epitope (12CAS) was purchased from Roche (Indianapolis, IN). Abs derived against IRF-3 and STAT1 were described previously (30, 33). Ab to IRF-7 was raised in rabbits against the 246–432 aa region of IRF-7. Anti-mouse or anti-rabbit secondary Abs conjugated to IRDye800 or 700 were obtained from Rockland Immunochemicals (Gilbertsville, PA). TNF-α was obtained from Invitrogen.
RNA was isolated from cells using SV Total RNA Isolation Kit (Promega), and cDNA was generated using random hexamer primers and SuperScriptII Reverse Transcriptase (Invitrogen) according to the manufacturer's protocol. RT-PCR was performed using the indicated primers and Taq polymerase (Invitrogen). Alternatively, quantitative real-time PCR was performed using the indicated primers at their optimal conditions as suggested by the manufacturer's instructions for the LightCycler-FastStart DNA Master SYBR Green I kit (Roche Molecular Biochemicals). Data was analyzed using the LightCycler software (Roche Molecular Biochemicals), and values were normalized to actin mRNA levels. Human primers used in the studies corresponded to: IFN-λ1 (37), IFN-α6 (38), IFN-α1/13 (38), actin (39), and pan–IFNα 5′-CACACAGGCTTCCAGGCATTC-3′ and 5′-TCTTCAGCACAAAGGACTCATCTG-3′; ISG54 (+1608) 5′-ATTCTATCACCAAGCCCGTGG-3′ and (+1370) 5′-TGGAGTCTGGAAGCCTCATCC-3′; IFN-α7 (+194) 5′-TCCTCCTCCGGGAATCTGAAT-3′ and (+106) 5′-AGGGCCTTGATACTCCTGG-3′; IFN-α14 (+315) 5′-TAGGAGGGTCTCATCCCAAGC-3′ and (+65) 5′-TGGGCTGTAATCTGTCTCAAAC-3′; IFN-β (+27) 5′-TGCTCTCCTGTTGTGCTTCTCCAC-3′ and (+243) 5′-ATAGATGGTC AATGCGGCGTCC-3′; IRF-7 (+1476) 5′-GATGTCGTCATAGAGGCT GTTGG-3′ and (+1343) 5′-TGGTCCTGGTGAAGCTGGAA-3′; IRF-1 (+397) 5′-TCTTAGCATCTCGGCTGGACTTC-3′ and (+190) 5′-CGATACAAAGCAGGGGAAAAGG-3′; and GAPDH (+1007) 5′-TACTCCTTGGAGGCCATGTG-3′ and (+528) 5′-CACAGTCCATGCCATCACTG-3′. Murine primers corresponded to: pan-murine IFN-α 5′-CCTGAGAGAGAAGAAACAC AGCC-3′ and 5′-TCTGCTCTGACCACYTCCCAG -3′ (40); murine IFN-λ2 (41); and murine Actin (42).
Immunoprecipitation and Western blot
For immunoprecipitation, cells were lysed in 50 mM HEPES (pH 7.2), 250 mM NaCl, 0.5% Nonidet P-40, 5 mM EDTA, 1 mM DTT, 1 mM NaF, 0.1 mM Na3VO4, and protease inhibitor mixture (Sigma-Aldrich, St. Louis, MO). Lysates were cleared by centrifugation for 5 min at 15,000 × g and reacted with Abs for 3 h at 4°C. Immunocomplexes were collected with Protein G-conjugated agarose (Invitrogen). For direct Western blot, cells were lysed in 50 mM Tris (pH 7.5), 400 mM NaCl, 0.5% Nonidet P-40, 5 mM EDTA, 10% glycerol, 50 mM NaF, 0.1 mM Na3VO4, and protease inhibitors. The lysates were cleared by centrifugation and directly added to SDS sample buffer. Proteins were separated on 8% SDS-PAGE and transferred to nitrocellulose membrane (Pierce Biotechnology, Rockford, IL). Membranes were reacted with indicated Abs, and images were detected using the Odyssey infrared imaging system (Li-COR Biosciences, Lincoln, NE). Alternatively, secondary anti-rabbit or anti-mouse Abs linked to HRP (Amersham/GE Healthcare, Piscataway, NJ) were used, and the membrane was incubated in ECL reagents and exposed to film.
Cells were seeded on coverslips, fixed in 4% paraformaldehyde, and either visualized directly for GFP fluorescence or permeabilized in 0.2% Triton X-100 before reaction with anti-Myc Ab. Secondary Abs were conjugated to rhodamine (Jackson ImmunoResearch Laboratories, West Grove, PA.). Coverslips were mounted in anti-fade solution (Vectashield; Vector Laboratories, Burlingame, CA). Images were captured with a Zeiss Axiovert 200 M digital deconvolution microscope or Zeiss LSM 510 META NLO two-photon laser scanning confocal microscope (Carl Zeiss).
Ubiquitination assay
The HA-Ub0R63K expression plasmid was transfected into a stable cell line expressing tetracycline repressor and responsive Myc-His–IRF-7 gene as described above. Doxycycline was added 24 h posttransfection with or without etoposide as indicated. Cells were lysed by sonication in 6 M guanidine HCl, 0.1 M Na2HPO4/NaH2PO4 (pH 8), and 10 mM imidazole. Lysates were incubated with Ni-NTA agarose beads (Qiagen, Chatsworth, CA), and proteins were eluted with SDS loading buffer and analyzed by Western blot (43).
Etoposide activation of IFN signaling
Cells respond to viral infection with the induction of ISGs, either directly by activation of the IRF-3 transcription factor or as an indirect response to autocrine IFN and STAT activation (15, 19, 30). Because the genes induced by IRF-3 or STATs can promote cellular apoptosis, we examined gene expression during the DNA damage response to identify shared proapoptotic-induced genes (24, 44–46). Etoposide, an anticancer drug, was used to initiate the DNA damage response because it inhibits the ability of topoisomerase II to religate cleaved DNA (47, 48). The inhibition results in an accumulation of double-stranded breaks in DNA, particularly during DNA replication and S/G2 phases of the cell cycle. The increase of dsDNA breaks with time leads to the DNA damage response and cell-cycle arrest or apoptosis. Following cell treatment with etoposide, we noted induction of several of the ISGs. The response of primary human monocytes was examined, and induction of a representative gene, ISG54/Ifit2, is shown in Fig. 1A. The effect of etoposide gradually increases the number of dsDNA breaks, and accordingly, the levels of ISG54 mRNA increased with time. To ensure this response was not specific to cell type, we evaluated the response of HeLa cells to etoposide. These cells also responded with induction of the ISG54 gene.
Induction of ISG54 mRNA and activation of STAT signaling in response to etoposide. A, Primary (1°) human monocytes were untreated or treated with etoposide for 0, 18, or 24 h. RNA was extracted, and real-time RT-PCR was performed with primers specific for ISG54 and quantified (left panel). HeLa cells were untreated or treated with etoposide for 24 h. RNA was extracted, and real-time RT-PCR was performed with primers specific for ISG54 and quantified (right panel). B, HeLa cells were transfected with STAT1-GFP or STAT2-GFP and untreated (−) or treated with etoposide for 24 h. Fluorescent images are shown. C, HeLa cells were treated with etoposide for the hours indicated, and cell lysates were evaluated for endogenous STAT1 (top panels) or STAT2 (bottom panels) tyrosine phosphorylation by Western blot. Results represent duplicate determinations in three independent experiments.
To access whether IFN production was responsible for ISG54 induction, we evaluated the activation of the STAT1 and STAT2 transcription factors in etoposide-treated cells. Binding of type I IFNs to specific receptors leads to tyrosine phosphorylation and nuclear accumulation of STAT1 and STAT2 (49). Cells expressing GFP-tagged STAT1 or STAT2 were examined for their response, and nuclear accumulation was evident by 24 h of etoposide treatment (Fig. 1B). In addition, the tyrosine phosphorylation of endogenous STAT1 and STAT2 was clearly evident (Fig. 1C).
These results suggested that etoposide treatment could induce the production and action of IFN, and accordingly, we evaluated the induction of IFN-α, IFN-β, and IFN-λ genes in primary human monocytes. There was little response of the IFN-β gene to DNA damage, although the gene was robustly induced in response to infection by NDV (Fig. 2A). In contrast, IFN-α mRNA expression was evaluated with pan-specific primers and clearly displayed an induction in response to etoposide (Fig. 2B). To determine the induction of IFN-α gene family subtypes, we tested the expression of individual genes. Various IFN-α genes tested were induced (Fig. 2C). We next tested expression of the newest family of IFN, IFN-λ. Specific induction of the IFN-λ1 gene clearly increased with etoposide treatment (Fig. 2D).
Specific IFN species are induced in response to etoposide. A, Primary human monocytes were untreated (−) or treated with etoposide for 20 h (20E) or 30 h (30E) or infected with NDV for 6 h (6V). mRNA levels were quantified by real-time RT-PCR corresponding to the IFN-β gene in response to etoposide or NDV. B, mRNA levels quantified by pan-specific primers to IFN-α from primary monocytes treated with etoposide as in A. C, mRNA levels were quantified to specific subspecies IFN-α1/α13, IFN-α6, IFN-α7, and IFN-α14 following 30-h etoposide treatment of primary monocytes. D, mRNA levels corresponding to the IFN-λ1 gene from primary monocytes treated with etoposide as in A. Values are means of duplicate determinations in two or three independent experiments.
Induction of ISG54, IFN-α, and IFN-λ in response to various DNA damaging agents
Etoposide elicits dsDNA breaks by forming an inactive ternary complex with topoisomerase II and inhibits the ability of the enzyme to religate cleaved DNA. Effectiveness can vary with proliferation of cell cultures, but 40 μg/ml etoposide was usually optimal for the IFN response (Supplemental Fig. 1). To determine whether IFN signaling is a general response to DNA damage or specific to etoposide, three diverse agents were tested: camptothecin, mitomycin C, and adriamycin. Camptothecin elicits ssDNA breaks by forming a ternary complex with topoisomerase I, mitomycin C is a DNA alkylating agent, and adriamycin causes DNA strand breaks by intercalation (50). Cells were treated with these agents and evaluated for ISG54 protein expression and mRNA levels of IFN-α and IFN-λ1 (Fig. 3). Despite the different mechanisms that elicit DNA damage by these agents, they all induced ISG54 and the IFN genes. The differences in fold induction may be a consequence of various peak response times due to different mechanisms of DNA damage. The results indicate that the production of IFN in etoposide-treated cells is a general DNA damage response and not one specific to a single drug or DNA insult.
Various DNA-damaging agents activate IFN signaling. THP-1 cells were untreated (−) or treated with 40 μg/ml etoposide (E), 5 μM camptothecin (C), 20 μg/ml mitomycin (M), or 20 μg/ml adriamycin (A) for 24 h. A, Cell lysates were prepared, and ISG54 protein levels were evaluated by Western blot. B, IFN-α mRNA induction was evaluated with pan-specific primers and real-time RT-PCR. C, IFN-λ1–specific mRNA expression was evaluated by real-time RT-PCR. Values are means of duplicate determinations in two independent experiments.
DNA damage activates IRF-1 and IRF-7 but not IRF-3
IRF-3 is expressed constitutively in cells, and in response to viral infection, it is a critical transcription factor for the induction of the IFN-β gene, a subset of IFN-α genes, and the direct induction of a subset of ISGs (15, 16, 51, 52). It exists in a latent state primarily in the cytoplasm and is modified by specific serine phosphorylation following viral infection. Phosphorylation promotes IRF-3 nuclear accumulation, DNA binding, and association with CBP. We evaluated these parameters for the activation of IRF-3 in response to DNA damage. Cells expressing IRF-3 tagged with GFP were treated with etoposide and visualized microscopically to access the cellular localization of IRF-3. IRF-3 remained primarily cytoplasmic and did not show evidence of nuclear accumulation (Fig. 4A). The phosphorylated forms of IRF-3 can be detected by their reduced migration during SDS-PAGE. This can be easily observed following viral infection, but it was not evident in response to DNA damage elicited with camptothecin or etoposide (Fig. 4B). The ability of activated IRF-3 to form complexes with CBP was evaluated by coimmunoprecipitation and Western blot. Although IRF-3 can be readily detected in immunocomplexes with CBP during viral infection, there was no evidence of association with CBP following etoposide treatment.
IRF-3 is not activated in response to DNA damage. A, HeLa cells were transfected with IRF-3–GFP and left untreated (−) or treated with etoposide for 24 h. Cellular localization was evaluated by fluorescence microscopy. Images represent random sampling from three independent experiments. B, Cells were untreated or infected with NDV for 6 or 12 h, treated with camptothecin (Camp) for 12 or 20 h, or treated with etoposide (Etop) for 20 or 40 h. Endogenous IRF-3 was detected in cell lysates by Western blot. C, CBP was immunoprecipitated (IP) from lysates of untreated cells or cells infected with NDV for 6 h or treated with camptothecin or etoposide for 36 and 50 h. Western blots (WB) were performed with Abs to IRF-3.
The IRF-7 transcription factor is a key regulator of the IFN-α genes in response to viral or bacterial infection (18, 29, 51, 53). It is expressed at low levels in lymphoid cells, but it is induced in all cell types by IFN and can function in a secondary wave of IFN production. We evaluated induction of the IRF-7 gene in response to DNA damage and found mRNA levels were clearly induced following etoposide (Fig. 5A). mRNA levels increased by >10-fold estimated by real-time PCR in 24 h (S. Brzostek-Racine and N. C. Reich, unpublished observations). IRF-7 can be activated in response to viral infection by serine phosphorylation. Although this modification may occur, it was not evident by a reduced migration in SDS-PAGE (S. Brzostek-Racine and N.C. Reich, unpublished observations) (54). Regulation of IRF-7 activity has also been demonstrated to occur by lysine 63-linked ubiquitination, and this modification was evaluated (55). A stable cell line was generated expressing a tetracycline/doxycycline-inducible IRF-7 gene tagged with the His and Myc epitopes. These cells were transfected with a gene encoding ubiquitin in which all of the lysines were mutated except lysine 63, and therefore, polyubiquitination could only occur via isopeptide linkage with lysine 63 (35). Cells were untreated or treated with etoposide, and IRF-7 protein was captured on nickel-charged resins and evaluated by Western blot (Fig. 5B). Following etoposide treatment, IRF-7 displayed slow migrating species indicative of its polyubiquitination and activation. Lysine 63-linked ubiquitin chains have been shown to regulate cellular processes including protein–protein interactions and to play a critical role in the DNA damage response (56).
Evidence for a role of IRF-7 in the IFN response to DNA damage. A, THP-1 cells were untreated (−) or treated with etoposide (+) for 24 h. IRF-7 mRNA induction was evaluated by RT-PCR and displayed on agarose gels. Faint band in untreated sample is nonspecific. mRNA levels of GAPDH are shown as controls. B, HT1080 stable cell line expressing tetracycline-inducible IRF-7–Myc-His was transfected with the HA-Ub0R63K ubiquitin (K63Ub), and doxycycline (Dox) was used to induce IRF-7 expression in the absence or presence of etoposide for 24 h. IRF-7 was collected on nickel-charged resins, and samples were analyzed by Western blot with Ab to IRF-7. Lower panel shows input before resin with anti-myc Ab. C, Expression of IRF-7–Myc-His was induced with doxycycline in the stable cell line in the absence or presence of etoposide for 15 h before immunostaining with Abs to Myc. Imaging analysis of three independent experiments indicate nuclear accumulation of IRF-7 at this time in >50% of the cells. D, Pan-specific primers were used to quantify IFN-α mRNA levels (top panel), and specific primers were used to assess IFN-λ1 mRNA levels (bottom panel) in the IRF-7–Myc-His–inducible cells by real-time PCR. Cells were untreated (−) or treated with etoposide (E), doxycycline (D), or doxycycline and etoposide (D/E) for 24 h. Values are means of duplicate determinations in two independent experiments.
In a latent state, IRF-7 resides primarily in the cytoplasm and accumulates in the nucleus following activation. The tetracycline-inducible cell line was used to evaluate IRF-7 localization by immunofluorescence during DNA damage (Fig. 5C). Following induction of IRF-7 expression, the protein was clearly cytoplasmic in untreated cells. However, IRF-7 accumulated in the nucleus following etoposide treatment, suggesting its activation. To evaluate the effects of inducible IRF-7 on IFN-α and IFN-λ1, their mRNA levels were measured (Fig. 5D). Cells were treated with etoposide, treated with tetracycline to induce IRF-7, or treated with both etoposide and tetracycline. Pan-specific primers were used to quantify expression of the IFN-α family. IFN-α was found to be induced in response to etoposide, but there was minimal effect of tetracycline-induced IRF-7 alone. However, etoposide and tetracycline potently increased etoposide-induced IFN-α expression. The results indicate modification of IRF-7 protein in response to DNA damage contributes to induction of the IFN-α genes. Expression of IFN-λ1 mRNA was similarly evaluated, and IRF-7 activation by etoposide also was found to contribute to induction of the IFN-λ1 gene.
Another member of the IRF family, IRF-1, has diverse roles in response to pathogens, development of the immune system, growth arrest, and apoptosis (15, 57, 58). The protein is constitutively nuclear but only expressed following induction by cytokines such as IFN. We evaluated IRF-1 mRNA expression and found mRNA levels increased by >12-fold estimated by real-time PCR following 24 h of etoposide treatment (Fig. 6A) (S. Brzostek-Racine and N.C. Reich, unpublished observations). To evaluate the possible role of IRF-1 in induction of the IFN genes during DNA damage, we tested its effect on expression of the IFN-λ1 promoter driving a luciferase reporter gene. The promoter of IFN-λ1 possesses IRF binding sites and a bona fide NF-κB site (21, 22). The IFN-λ1 promoter activity was induced following etoposide treatment or by cotransfection with a plasmid encoding IRF-1 (Fig. 6B). Combined IRF-1 expression with etoposide induced significant expression of the IFN-λ1 promoter. IRF-1 could also be demonstrated to bind to a site in the IFN-λ1 promoter, suggesting direct action on the IFN-λ1 gene (Supplemental Fig. 2).
Activation and inhibition of IRF and IFN genes. A, THP-1 cells were untreated (−) or treated with etoposide (+) for 24 h. IRF-1 mRNA levels were evaluated by RT-PCR and displayed on agarose gels. mRNA levels of actin are shown as controls. B, The IFN-λ1 luciferase reporter was expressed in HeLa cells untreated or treated with etoposide (Etop) for 24 h. Empty vector (c) or IRF-1 expression plasmid (IRF1) were cotransfected where indicated with or without etoposide treatment and luciferase activity was measured. C, Effects of the ATM inhibitor AZ12622702 (AZ) on IFN-α and IFN-λ gene expression. HeLa cells were untreated or treated with AZ for 1 h followed by etoposide (E) for 24 h as indicated. Real-time PCR was used to quantify IFN-α (left panel) or IFN-λ (right panel) mRNA expression. D, Effects of IKKβ inhibitors BAY117085 (BAY) or ML120B (ML120) on IRF-1 and IRF-7 gene expression. HeLa cells were untreated or treated with the inhibitors for 1 h followed by etoposide for 5 h as indicated. Real-time PCR was used to quantify endogenous IRF-1 (left panel) or IRF-7 (right panel) mRNA expression. Quantitative results are means of duplicate determinations in three independent experiments.
dsDNA breaks that occur in response to etoposide lead to the recruitment and activation of the ATM kinase. To evaluate the potential role of ATM in the induction of IFN gene expression, we tested the effects of a pharmacological inhibitor, AZ12622702. Treatment of cells with the ATM inhibitor decreased the ability of etoposide to induce the IFN-λ1 gene and the IFN-α genes (Fig. 6C). The results indicate ATM is a significant signaling kinase upstream of IFN gene induction, and the residual response may be due to other PI3K-related kinases that are activated during DNA damage.
Role of NF-κB in the response of IFN genes to DNA damage
]IRF-1 and IRF-7 are both induced in response to IFN signaling and to DNA damage. Studies have also indicated that the promoters of both of these genes can be activated by the NF-κB transcription factor (59–61). Because NF-κB activation is a well-characterized response to DNA damage and ATM activity, we determined whether induction of IRF-1 and IRF-7 by etoposide is a direct response to NF-κB (11, 62). Inactive NF-κB is localized in the cytoplasm due to binding IκB (63). IκB is released following its phosphorylation by an IKK complex composed of IKKα, IKKβ, and NEMO/IKKγ. To determine the role of NF-κB in induction of IRF-1 and IRF-7 by DNA damage, we tested the effect of several specific IKKβ inhibitors (Fig. 6D). Cells were treated with etoposide in the absence or presence of BAY117085 or ML120B, and real-time PCR was used to quantify the endogenous expression of IRF-1 and IRF-7 mRNAs. Both inhibitors effectively blocked the induction of IRF-1 and IRF-7 expression, indicating activation of NF-κB in response to DNA damage is necessary to induce these genes.
A major species of NF-κB is the p50 and p65/RelA heterodimer (63). Because serine phosphorylation of the C terminus of p65 has been shown to increase its transcriptional activity and stability, we evaluated the phosphorylation of p65 following etoposide treatment (64). Etoposide stimulated the serine 536 phosphorylation of p65, indicating the NF-κB is transcriptionally active (Fig. 7A). The activation of NF-κB can also be detected by its ability to accumulate in the nucleus. For this reason, we performed immunofluorescence staining of the endogenous p65 subunit before and after etoposide treatment. Cells treated with etoposide clearly displayed p65 nuclear accumulation (Fig. 7B). These properties accompanied the ability of NF-κB to bind a consensus DNA site in EMSAs (S. Brzostek-Racine and N.C. Reich, unpublished observations). The data support NF-κB activation in our system of DNA damage, as reported previously (11).
NF-κB is activated and required for IFN gene induction in response to etoposide. A, HeLa cells were treated with etoposide for 15 h or 5 ng/ml TNF-α for 1 h, and p65 was immunoprecipitated (IP) from lysates. Specific Ab to p65 phospho-serine 536 was used for the Western blot (WB). Lower panel displays Western blot with Ab to p65. B, HeLa cells were untreated or treated with etoposide for 2 h before immunostaining with Abs to p65. C, HeLa cells were untreated or treated with etoposide (E) in absence or presence of ML120B. Left panel, Real-time PCR was used with pan-specific primers to quantify the endogenous levels of IFN-α mRNA. Right panel, Real-time PCR was used to quantify the endogenous levels of IFN-λ1 mRNA. D, Left panel, The IFN-λ1 luciferase reporter plasmid was cotransfected with empty vector or with a plasmid encoding the dominant-negative IκBS32A/S36A (IκBSS/AA) gene. Cells were untreated or treated with etoposide for 24 h prior to the luciferase assays. Right panel, The IFN-λ1 luciferase reporter plasmid was cotransfected with empty vector (c) or with a plasmid encoding the p65/RelA gene. Cells were untreated or treated with etoposide for 24 h prior to luciferase assays. Quantitative results are means of duplicate determinations in three independent experiments.
To evaluate the role of NF-κB in the induction of IFN genes by etoposide, we tested the effects of IKKβ inhibition. Cells were treated with etoposide in the absence or presence of the IKKβ inhibitor ML120B, and IFN-α mRNA levels were quantified by real-time RT-PCR (Fig. 7C). Inhibition of IKKβ was found to block expression of the IFN-α genes, indicating that NF-κB is required for induction. This is a significant finding because the IFN-α genes are not directly regulated by NF-κB and do not possess NF-κB binding sites, although they do possess IRF binding sites (51).
Induction of the IFN-λ1 gene by DNA damage was also was found to be dependent on NF-κB. Real-time RT-PCR was used to quantify IFN-λ1 mRNA levels following etoposide in the presence or absence of ML120B. Expression of IFN-λ1 was blocked with the inhibition of IKKβ and NF-κB. The effect of NF-κB inhibition could also be demonstrated with the IFN-λ1 promoter driving the luciferase gene. The IFN-λ1 promoter reporter was expressed alone or with a dominant-negative IκB mutant that lacks serine target phosphorylation sites (S32A/S36A) (34). Etoposide stimulated the expression of the IFN-λ1 luciferase reporter, but this induction was completely inhibited with coexpression of the NF-κB repressor IκB (S32A/S36A) (Fig. 7D).
Because the promoter of the IFN-λ1 gene contains an NF-κB binding site, the more direct question is whether NF-κB in the absence of viral infection or DNA damage can induce the IFN-λ1 gene (21). To determine whether NF-κB regulates induction of the IFN-λ1 gene, we tested the response of the IFN-λ1 promoter reporter to expression of p65/RelA, a potent activator of NF-κB target sites. The IFN-λ1 gene was induced directly by p65 coexpression, indicating NF-κB is sufficient to induce the human IFN-λ1 gene independent of viral infection or DNA damage (Fig. 7D). This result has obvious implications for the involvement of IFN action in the many signaling pathways that activate NF-κB.
Response of murine embryo fibroblasts
Our studies with human primary cells or established human cell lines provide clear evidence that the IFN-λ and IFN-α genes are induced in response to DNA damage by etoposide and that NF-κB is requisite for the induction. Although murine cells may not accurately reflect the human response, the murine system affords the ability to test cells from animals with specific gene knockouts. For this reason, we obtained MEFs from wild-type (wt) or gene knockout animals and tested their response to DNA damage. Wild-type MEFs or MEFs that lack NEMO were treated with etoposide, and IFN gene induction was evaluated by RT-PCR (Fig. 8). Because murine IFN-λ1 is a pseudogene, we assayed expression of murine IFN-λ2. Results showed etoposide treatment induced the genes encoding IFN-λ2 and IFN-α in wt cells, but not in NEMO knockout cells. The results are in accordance with our studies in human cells that demonstrated a requirement of NF-κB activation. In addition, MEFs from IRF3 knockout animals treated with etoposide induced the IFN-λ2 and IFN-α genes, supporting our studies with human cells that showed IRF3 did not play a major role in the DNA damage response. Analyses of MEFs that lack the IRF-1 gene indicated it was critical for IFN-λ2 gene expression, but not for IFN-α expression, whereas MEFs that lack the IRF-7 gene had a profound defect in the induction of both IFN-λ2 and IFN-α genes in response to etoposide. The promoters of the murine IFN-λ genes are not well characterized, and therefore, they may respond differently from the human IFN-λ genes during the DNA damage response. The levels of IFN induction in these spontaneously immortalized MEFs were modest but reproducible following etoposide treatment.
Effect of etoposide on MEFs. MEFs isolated from wt mice or mice with targeted gene knockouts in NEMO (NEMO−/−), IRF-3 (IRF3−/−), IRF-1 (IRF1−/−), or IRF-7 (IRF7−/−) were untreated or treated with etoposide for 24 h. Real-time RT-PCR was used to quantify endogenous levels of murine IFN-λ2 (A) or pan–IFN-α (B). Quantitative results are means of duplicate determinations in three independent experiments.
Expression of human IRF and IFN genes with time during the DNA damage response
Our studies with human cells indicate that NF-κB activated by DNA damage stimulates induction of the IFN-λ1, IRF-1, and IRF-7 genes. To determine the time course of expression of these genes, human THP-1 cells were treated with etoposide, and real-time PCR was used to quantify IRF and IFN mRNA levels. IRF-1 and IRF-7 mRNA levels displayed an initial peak of expression at 4 h of etoposide treatment and reached steady-state levels by ∼12 h (Fig. 9A). Expression of the IFN-λ1 gene showed a small increase at 4 h of etoposide treatment and peaked at 15 h with a kinetic profile similar to that of the IRFs. Expression of IFN-α mRNA trailed that of IFN-λ1 by 3 to 4 h, possibly indicating a greater dependency on IRF-7 induction and activation (Fig. 9B).
Time course of IRF and IFN mRNA expression in the response to etoposide treatment. THP-1 cells were treated with etoposide, and RNA was isolated from cells during a 24-h period at the times indicated. Real-time RT-PCR with specific primers was used to quantify IRF-1 and IRF-7 mRNA (A) or IFN-α and IFN-λ1 mRNA (B). Results are means of two experiments performed in duplicate. C, Conceptual model of dsDNA break response signaling to IFN gene expression. ATM phosphorylation of NEMO promotes its ability to bind and activate the IKK complexes. IKKs phosphorylate IκB, releasing NF-κB to activate gene targets IFN-λ1, IRF-1, and IRF-7. The IRFs induce IFN-α genes and enhance IFN-λ1 expression.
The DNA damage response rapidly engages multimeric protein complexes to repair DNA and activate transcriptional programs that regulate cell-cycle checkpoints and cell survival (6, 8, 65, 66). The recruitment of ATM, ATR, and DNA-PK to DNA breaks initiates phosphorylation and ubiquitination events that lead to specific transcription factor activation and gene expression. ATM is activated primarily in response to dsDNA breaks, followed by ATR and DNA-PK in response to DNA single strands and ends generated during break resolution. One of the known substrates of ATM is NEMO, and phosphorylation promotes its ubiquitination, nuclear export, and activation of IKK complexes (62, 67, 68) (Fig. 9C). IKK phosphorylation of IκB leads to release of NF-κB dimers and their ability to translocate to the nucleus and bind DNA targets. Our studies demonstrate that NF-κB is sufficient to induce the human IFN-λ1 gene during the response to DNA damage. The promoter of the human IFN-λ1 gene has a bona fide NF-κB binding site as well as an IRF binding site (21). NF-κB also induces the IRF-1 and IRF-7 genes that can influence expression of the IFN-α and IFN-λ genes. The induced IFNs can additionally amplify expression of the IRFs.
These findings add a new dimension to the complexity of the DNA damage response. ATM appears to be primarily accountable for initial signal pathways that lead to human IFN gene expression by etoposide. Inhibition of ATM significantly reduces the induction of IFN-α and IFN-λ1 genes (Fig. 6). The ATM deficiency responsible for the development of ataxia-telangiectasia results in an array of clinical manifestations including cerebellar ataxia, oculocutaneous telangiectasia, immunodeficiency, and susceptibility to cancer (69, 70). The lack of IFN production in response to DNA damage that occurs through physiological processes in ataxia-telangiectasia speculatively may contribute to the immunodeficiency and tumor formation in the disease. Results of a few studies have suggested a potential role of IFN signaling during DNA damage (71–74). One study observed ISG expression correlated with resistance to radiation therapy (74), and another study reported that STAT1 facilitated cell-cycle checkpoint following DNA damage (72). IFN pathways not only stimulate Janus kinases and STAT factors but also elicit a broad range of effects on transcription and translation. The contribution of IFN signaling in the response to DNA breaks may have multifaceted consequences.
The novel observation of the convergence of the DNA damage response with IFN signaling stimulates speculation as to the possible function of IFN in reaction to genotoxic stress. In our experimental system, the addition of IFN did not block the apoptotic effects of etoposide or significantly contribute to cell death (S. Brzostek-Racine and N.C. Reich, unpublished observations). But IFNs are well characterized for their ability to inhibit viral infection, and this may reflect the evolutionary link. Many viral infections are known to stimulate DNA damage response pathways. Viruses like HIV have RNA genomes but integrate viral DNA into the host genome, creating DNA strand breaks (4). Viruses with DNA genomes can generate ss- and dsDNA breaks during lytic replication. EBV, HSV1, and adenovirus are a few examples of DNA viruses that have been documented to activate a DNA damage response (4, 75, 76). More significantly, some of these viruses have evolved mechanisms to inhibit activation or downstream function of the DNA damage response (77–79). Viruses may inhibit this pathway not only to block cell cycle arrest and apoptosis, but also to block the antiviral functions of IFNs that are produced by DNA damage.
Etoposide and IFN both have been used clinically for years for their antitumorigenic effects. IFNs produced in response to DNA damage may contribute to the antitumorigenic effects of etoposide. IFNs are recognized for their ability to cause growth arrest and/or apoptosis in neoplastic cells, although they can stimulate proliferation of healthy cells (24, 25, 46, 80–83). They also have vital immunoregulatory functions that include direct and indirect effects on activation of NK cells, macrophages, dendritic cells, T cells, and B cells (84, 85). The antiproliferative effects of IFN and the potential enhanced clearance of tumor cells may play a role in the in vivo DNA damage response pathway.
The profile of IFN gene expression in response to DNA damage in human cells is distinct from that induced by viral infection. The existence of multiple IFN genes with distinct promoter elements appears to have evolved as a response to different cellular stresses. A significant finding of our study is the ability of NF-κB to stimulate the expression of IFN genes in the absence of viral infection. NF-κB is required for the induction of IRF-1, IRF-7, IFN-λ, and IFN-α in response to etoposide. Although the promoter of the human IFN-λ1 gene possesses both an NF-κB binding site and an IRF binding site, the promoters of the IFN-α genes possess only IRF binding sites. Results with the knockout MEFs indicate a critical function of IRF-7 activated in response to etoposide for induction of both murine IFN-λ2 and IFN-α genes. Future studies are needed to provide additional insight on the impact of IRF-1 and IRF-7 on the IFN genes during the DNA damage response and whether there are significant mechanistic differences in the human versus murine response. NF-κB is activated not only by DNA damage but also by a wide array of cellular stimuli, and it plays a major role in inflammation, immunity, cell survival, and cancer (63). The intimate link of NF-κB to IFN gene induction during the DNA damage response may reflect a potential role of IFN in other biological responses to NF-κB in the absence of infections. Our study adds a significant finding of IFN signaling to the complexity of pathways that are orchestrated by the response to genotoxic stress.
The authors have no financial conflicts of interest.
We thank all of the members of the Reich laboratory for support. We also thank Dr. Martha Furie, Gregory Sabino, and Indralatha Jayatilaka for helpfulness in preparation of primary monocytes.
This work was supported by National Institute of Allergy and Infectious Diseases Grants R21AI067885 and PO1AI0555621 and National Institutes of Health Grant T32GM008444.
Abbreviations used in this article:
ataxia-telangiectasia mutated
ataxia-telangiectasia mutated and Rad3 related
CREB-binding protein
polyhistidine
IκB kinase
IFN regulatory factor
IFN-stimulated gene
Murine embryo fibroblast
NDV
NF-κB essential modulator
wild-type.
Received January 5, 2011.
Accepted September 12, 2011.
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Vol. 187, Issue 10
You are going to email the following The DNA Damage Response Induces IFN
Sabrina Brzostek-Racine, Chris Gordon, Sarah Van Scoy, Nancy C. Reich
The Journal of Immunology November 15, 2011, 187 (10) 5336-5345; DOI: 10.4049/jimmunol.1100040


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Accounting & Finance Investment Management
BSc Economics and Management
經濟及管理學理學士
CEF Reimbursable Course (selected modules only)
University of London (Academic Direction: London School of Economics and Political Science)
MS025A
QF Level: 5 QR Registration No.: 13/000312/L5 QR Registration Validity Period: 01 Sep 2012 - 31 Aug 2026
Continuing Education Fund Reimbursable Course
Some modules of this course have been included in the list of reimbursable courses under the Continuing Education Fund.
04 Sep 2023 (Mon)
2 years to 6 (Max) years
Admission: 3761-1122
Tuition Course: 2867-8319
lse@hkuspace.hku.hk
The degree is awarded by the University of London and the programme is...
This degree programme allows students to achieve the knowledge of 'effective management' to understand the operation of organisations by exploring different disciplines; economics, finance, organisation theory, mathematics and statistics. The programme is also suitable for people who are interested in general management, further studies in management or related disciplines, as it provides an in-depth study of the both economics and general management options.
Programme Objectives
This programme aims to:
provide a high quality education in Economics and Management for intellectually able students from a wide range of backgrounds;
give students a sound grasp of the principles of economic analysis to the intermediate level;
help students understand the social science basis and inter-disciplinary nature of Management;
give students a knowledge of the workings of the economy;
enable students to concentrate on specialist areas within the subjects;
encourage students to develop intellectual flexibility and powers of rigorous analysis, by placing a premium on developing minds and on bringing forward their analytical reasoning ability;
develop in students the ability to apply their knowledge to the solution of theoretical and applied problems in economics and management;
provide a basis for further study and for employment, including careers in relevant fields;
prepare students for graduate study in economics and/or management.
Intended Learning Outcomes (ILOs) of the Programme
On completion of the programme, students should be able to
A. Knowledge and understanding
understand of the fundamental aspects of economic analysis. (modules 1, 6, 7, and 9, plus modules 10 to 12 elective a )
analyse issues from the perspective of a number of disciplines. (modules 1 to 12)
think in a critical manner, to formulate and develop arguments both on management and economic issues in a logical manner. (modules 4, 5, 6b, and 9, plus modules 10 to 12 selected from electives a to e and g)
demonstrate skills in applying economic reasoning to the analysis of a wide range of policy issues. (modules 1, 6, and 7, plus modules 10 to 12 elective d)
B. Intellectual and cognitive skills
locate, extract, analyse and draw reasoned conclusions from multiple sources (including electronic sources) of appropriate literature and relevant data and acknowledge and reference these sources appropriately; (modules 1 to 12)
critically analyse arguments; (modules 1, 2b, and 3 to 12)
apply theory and theoretical models in practical situations; (modules 3, 4, 5, 6b, and 8, plus modules 10 to12 selected from electives f and h)
assimilate and lucidly evaluate alternative views. (modules 1, 2b, and 3 to 12)
C. Transferable skills
communicate effectively in a written context; (modules 1, 2b, and 3 to 12)
use information technology (spread-sheets, word-processing and online databases) appropriately; (modules 3, and 8, plus modules 10 to 12 selected from electives d, f to i)
organise information, and assimilate and evaluate competing arguments; (modules 1, 2b, and 3 to 12)
manage his or her own learning, including working effectively to deadlines; (modules 1 to 12)
be open minded and have a capacity to handle ideas and scrutinise information in critical, evaluative and analytical ways. (modules 1 to 12)
Programme Structure (Study at HKU SPACE)
The entire programme consists of 12 modules (30 credits each).
Upon completion of 12 modules (360 credits), students will be awarded the Bachelor of Science in Economics and Management by the University of London. No other exit awards are accredited by HKCAAVQ.
A. Programme Structure (Standard Route)
The below structure is for students first registered in 2023/24, for students who registered in 2022/23 or earlier, the programme structure should be followed by the programme regulation on the University of London Website.
All programmes in Standard Route consist of 12 courses.
Minimum Duration: 3 years part time*
* Students who gain 3 APL may complete the programme in 2 years, subject to individual case.
For students first registering for a degree in 2023/24
To be considered for an honours qualification, you must attempt 12 full courses of the study programme, and pass a minimum of 11 full courses.
To be considered for a pass without honours, you must attempt 10 full courses of the study programme, and pass 10 full courses.
No other exit awards are accredited by HKCAAVQ.
For details, please refer to the University of London programme regulations: http://www.londoninternational.ac.uk/
The Course Structure of BSc Economics and Management**
Level 100 courses
1. Introduction to Economics
2. Mathematics 1 (Half course) and Statistics 1 (Half course)
3. Principles of Accounting
4. Business and Management in a Global Context
Level 200 / 300 courses
5. Core Management Concepts
6. Choose ONE from below:
7. Macroeconomics
8. Asset Pricing and Financial Markets
9. Monetary Economics
10-12. Choose THREE from below
a. Strategy*
b. Organisation Theory: an interdisciplinary approach*
c. Human Resource Management*
d. Marketing Management*^
e. The Law of Business Organisations*
f. Management and Innovation of e-business*
g. Management Accounting
h. Information Systems Management
i. Principles of Corporate Finance
j. Principles of Banking and Finance
* Students should take at least one module.
^ The course was named "Principles of Marketing" before.
** Subject to change and approval.
The above programme structure is intended to be used as a guide by prospective students. Students must also follow the programme structure presented by the University of London International Programmes Programme Specification and Regulations.
HKU SPACE may not offer all courses in each year.
B. Programme Structure (Graduate Entry Route)
Recognized degree holders can study the BSc Programmes through Graduate Entry Route.
The programme in Graduate Entry Route consists of 9 courses.
Students who are admitted through the Graduate Entry Route (Refer to the Entry Requirements) will have to take courses 1,2,4,5,6,7,8,9, and one course from a to i under courses 10 to 12 .
For the Graduate Entry Route degree programmes, you must successfully pass all nine courses to be eligible for the award of degree.
Students registered for BSc Economics and Management through the Graduate Entry Route may not be able to finish in 2 years. Subject to individual case.
The course syllabus is available in the University of London prospectus:http://www.londoninternational.ac.uk/
Degrees are awarded with First Class Honours, Second Class Honours, Third Class Honours or Pass.
The above programme structure is intended for use by prospective students for the University of London BSc Programmes at HKU SPACE and is subjected to the final approval.
Course Fee#
Total fees: Around HK$74,785 + GBP6,821 (2022/23 enrolment) (3-year part-time study)
Year 1:
Application Fee*: GBP107
Initial Registration Fee*: GBP1,214
HKU SPACE Tuition Course Fee (4 full courses): HK$5,400 x 4 = HK$21,600
Exam fee (4 full courses and to be adjusted in Dec 2023):
GBP1,464* + HK$3,815 (enrolled in 2017-18 or after)/GBP1,208* + HK$3,815 (enrolled in 2016-17 or earlier)
Total fee for Year 1: GBP2,785* + HK$25,415 (enrolled in 2017-18 or after)/
GBP2,529* + HK$25,415 (enrolled in 2016-17 or earlier)
Continuing Registration Fee*: GBP554 (enrolled in 2017-18 or after)/GBP514 (enrolled in 2016-17 or earlier)
Exam fee (4 full course and to be adjusted in Dec 2023):
Total fee for Year 3: GBP2,018* + HK$24,685 (enrolled in 2017-18 or after)
* All the fees paid in GBP are subject to the exchange rate.
# The fees are for 2022/23 academic year and for reference only. All the fees are reviewed annually and are subject to change without prior notice.
Assessment will be done by examination. Each course is assessed by one written examination determined by the University of London. All examinations are held once a year in May/June.
Standard Route (To obtain the FIRST Degree)
Minimum: 3 years*
Maximum: 6 years
Graduate Route (To obtain the SECOND Degree)
Minimum: 2 years^
^ Students who admitted through Graduate Route may not be able to complete the programme in 2 years, subject to individual case.
Each HKU SPACE tuition course comprises a total of 54 teaching hours.
Classes will normally be held on weekday evenings and weekends which may include Sundays & public holidays.
Admiralty Learning Centre
United Learning Centre
Fortress Tower Learning Centre
Non-Local Higher and Professional Education (Regulation) Ordinance
This is an exempted course under the Non-local Higher and Professional Education (Regulation) Ordinance. It is a matter of discretion for individual employers to recognise any qualification to which this course may lead.
Introduction to Economics
This course aims to introduce students to an understanding of the domain of economics as a social science; the main analytical tools and reasoning used in economic analysis; the main conclusions derived from economic analysis. It will also develop students' understanding of their organizational and policy implications and enable them to participate in debates on economic matters.
Mathematics 1 (half course)
The course aims to enable students to acquire skills in the methods of calculus (including multivariate calculus) and linear algebra, as required for their use in economics-based subjects. It also prepares students for further study in courses of mathematics and/or related disciplines.
Statistics 1 (half course)
The emphasis of the course is on the application of statistical methods in management, economics and the social sciences. Attention will focus on the interpretation of tables and results and the appropriate way to approach statistical problems. Treatment is at an elementary mathematical level. Ideas of probability, inference and multivariate analysis are introduced and are further developed in the half course 04b Statistics 2.
This course aims to: introduce students to the principles underlying accounting; enable students to apply, interpret and explain key accounting techniques; provide a broad understanding of the theory and practice of financial accounting and management accounting.
Business and Management in a Global Context
This module aims to: give students a research-based grounding in the shifting context of, and changing priorities in, international business including globalizing trends, formal, and informal institutions, the political, economic, social, technological and legal issues and the resultant diversity of international business. Specifically takes into account the impacts of the 2020-21 pandemic and economics crises; prepare students to be able to discuss cultural, ethical and social issues for international business and suggest policies of corporate social responsibility and sustainable business practices; provide an introduction to and develop students' ability to assess international trade and investment, multilateral organisations and regional integration and the global financial system; prepare students to be able to discuss how firms develop international business strategies, enter markets and alliances, and operate and manage on the global stage; give insight through frameworks, studies and examples of how businesses manage marketing and research & development, organisational structure and architecture, sourcing and the supply chain, exchange rates, information systems and human resources, in different parts of the globe, globally, regionally and domestically; prepare students to understand and assess project and change management practices in an international context; provide an introduction to the major emerging digital technologies, and how to manage these into operation and become digital businesses internationally; Overall, give students insight into the contexts of international business and enable students to work within these contexts to make judgements on strategizing and managing operations in the global economy. Also, to help students understand and assess the future trends and challenges for international business.
Core Management Concepts
The aims of the course are to: give students a thorough grounding in the key management sub‐disciplines; provide an overview of the development of these disciplines ; illustrate the disciplinary anchors of these disciplines in sociology, psychology and economics.
The course aims to: deepen the understanding of the basic theory of optimization by economic agents and the efficiency of the resulting outcome for the market as a whole; introduce students to the analysis of strategic interaction as well as interaction under asymmetric information; clarify the role of economic policies as tools to improve efficiency in the presence of market failures; promote the ability to think in a structured framework, and clarify the importance of formal arguments; demonstrate the art of modelling which requires simplifying a problem by identifying the key elements without oversimplifying the issue.
The course aims to: enable students to approach managerial decision problems using economic reasoning; present business practice topics using an analytical approach, using equations and numerical insight.
The course aims to bring the students up to date with modern developments in macroeconomics and to help them analyse the macroeconomic issues of the day.
Asset Pricing and Financial Markets
The course aims to provide students with a thorough grounding in asset pricing; develop students' skills in applying pricing methods to realistic scenarios; provide a critical overview of the research on financial market efficiency; allow students to develop an understanding of how securities markets operate.
The aims of the course are to develop students' understanding of the theories that relate to the existence of money, explaining why it is demanded by individuals and used in the trading process; develop students' understanding of the monetary transmission mechanism, whereby decisions made by the monetary authorities concerning money supplies or interest rates can have real effects on the economy; develop a number of macroeconomic models through which monetary policy can be evaluated. Such models will include both Classical and Keynesian schools of thought and will consider why monetary policy matters and when monetary policy decisions may be impotent; develop understanding of the uncertainties policy-makers face and how policy makers may deal with these.
The course aims to enable students to use the analytical tools of game theory and industrial economics to study key strategic problems in corporate decision making.
Organisation Theory: an interdisciplinary approach
This course explores the ways in which organisations can be designed (by management or others) in order to achieve their objectives. It exposes students to the ways in which economists, psychologists and sociologists address this and related issues. It is, therefore, an interdisciplinary course comparing and contrasting the contribution of the core social science disciplines to the study of organisations. The course draws upon both theoretical and empirical materials and students will be encouraged to draw upon their own local knowledge in pursuing their studies. There will be an emphasis (though not exclusive) on organisations, set up with an economics objective.
This course aims to: give students an introduce to the key elements of human resource management (HRM); demonstrate how the social sciences can assist in understanding the management of human resources; and to examine and evaluate human resource policies and practices of organizations; help students examine the different theories which try to explain the relationship between HRM and organisational performance; develop students' ability to analyse and critically evaluate HR policies and practices.
Marketing Management^
This course aims to introduce students to the fundamental principles of marketing; give students a broad understanding of consumers and the marketing behaviour of firms; explores the relevance of other academic disciplines to marketing; encourage students to question the limitations of marketing management and to suggest ways of overcoming its many problems; develop students' practical skills by applying learned theories to real-world organisational problems.
The Law of Business Organisations
This course aims to: develop in the students an understanding of the core principles of the laws that regulate business organisations; develop the independent and individual skills needed for students to take their knowledge further, and to apply it, for example, in practice, in industry, or in an academic environment; inspire and to stimulate endeavor.
Management and Innovation of e-business
The objectives of the course are to explain the growth of e-business to date and introduce the most relevant e-business models, using theories from business, management and the social sciences; examine the interaction between technological trends and the business and social context of e-business; identify innovations within the domain of e-business by presenting cases of the innovative use of e-business and network technologies; present relevant theories from business, management and the social sciences that help to explain the development and growth of e-business; discuss different e-business (business) models and strategies, including global supply chain management; electronic markets; shared economy and digital marketing; introduce the notion of technologically mediated organisational forms, and discuss their business implications.
This course is designed to equip students with knowledge of the concepts and the ability to apply techniques of management accounting, in order to be able to contribute to the success of an organisation. Modern management accounting is concerned with decision making, planning and control. It supports decision makers in their daily and strategic decisions, contributes to short- and long-term planning and produces information for control, performance measurement and management. This may also include nonfinancial information in management accounting reports, where it is used in combination with financial information to construct broader pictures of the contribution that different processes, procedures, functions, activities and divisions make to the strategic achievements of a firm.
Principles of Corporate Finance
This course provides a theoretical framework used to address issues in project appraisal and financing, payout policy, capital structure, mergers and acquisitions, equity offerings, and risk management. It provides students with the tools required for further studies in financial intermediation and investments.
Principles of Banking and Finance
The specific aims of this course are to: provide institutional features of financial systems; identify key issues and problems arising in banking and finance; introduce the key economic concepts required to analyse these key issues and problems; illustrate how these economic principles can be applied to address the key issues identified; show how the institutional features address the key issues identified.
Timetable for BSc Programmes Tuition Courses
(2022/23)*
Week A (Start date: 5 Sep 2022)
Organisation Theory: an
interdiscipliary approach Principles of Accounting Strategy The Law of Business Organisations Asset Pricing and Financial Market Some lessons will be arranged on Sat/Sun.
(1 or 2 consecutive lessons per day)
Auditing and Assurance Management and Innovation of e-business Mathematics 1 and Statistics 1
Monetary Economics Human Resource Management
Week B (Start date: 12 Sep 2022)
Macroeconomics Microeconomics Business and Management in a Global Context Principles of Banking and Finance Marketing Management^ Some lessons will be arranged on Sat/Sun.
Introduction to Economics Managerial Economics Core Management Concepts Financial Intermediation Management Accounting
Financial Reporting Principles of Corporate Finance Accounting: Markets and Organisations^^ Investment Management
* The timetable is designed for 2022/23 academic year and is for reference only. The timetable for 2023/24 will be updated later.
^^ The course was named "Accounting Theory" before.
Each HKU SPACE tuition course comprises a total of 54 teaching hours (16 lessons).
Lessons will be arranged on weekdays from 7:00 - 10:20 pm (3 hours and 20 minutes) & Saturday (s) / Sunday (s) (3-7 hours).
Classes will be held in Admiralty or other HKUSPACE learning centres, subject to availability.
Courses in week A & B will be held every 2 week alternatively.
Students are not allowed to take more than one course from each week on the same day as class crush will be occurred, and no refund will be made.
Make up class(es) will be held on Sunday(s) and/or Public Holiday(s).
Please check the programme structure and the prerequisites before enrolment. Course Transfer fee HK$120 will be charged if students would like to transfer from an enrolled course to another one (14 working days to process the transfer).
The School reserves the right to revise the above timetable without prior notice and cancel any tuition courses due to insufficient enrolment numbers.
HKU SPACE Vaccine Pass
To minimize the risk of COVID-19 transmission in learning centres and offices and to keep the HKU SPACE community safe, access to the HKU campus or HKU SPACE premises will need to comply with the tightened measures in line with the University. Please refer to details on implementation on our Website/Learner Portal and please do note that there may be further updates from time to time to take account of possible changes in the Government guidelines and University arrangements.
Class arrangement during the COVID-19
The COVID-19 situation may still be fluid and constantly affect class arrangements in the coming months. The health and safety of our students will always be our top priority. To ensure that students' academic progress is not affected, the School may substitute face-to-face classes with online teaching if necessary in the event that face-to-face classes cannot be held. Our respective Programme Teams will contact the students concerned with details of such arrangements as necessary.
Standard route*
Applicants shall:
2 HKDSE Category A subjects at Level 3 (including English Language) and 3 HKDSE Category A subjects at Level 2 (including Mathematics); or
(a) 2 GCE 'A' Level Grade E + 3 GCE 'O' Level Grade C (including English Language & Mathematics); or
(b) 2 HKAL Grade E + HKCEE English Language at Grade C & 2 subjects at Grade E in HKCEE (including Mathematics); or
International Baccalaureate (IB) Diploma including Mathematics / Mathematical Studies at Grade 3 (Higher Level / Standard Level); or
An Associate Degree / a Higher Diploma from HKU SPACE or other recognised awarding institutions.#
* Subject to change.
# Course specific requirements such as competence in English Language and Mathematics may be required.
Note: There are many other qualifications, both from Hong Kong and overseas, that the University of London accepts as satisfying the entrance requirements for the degree programmes. For details, please contact the HKU SPACE University of London Programmes (Admissions Office) at 3761-1122 or email londonu@hkuspace.hku.hk
Students transfer to the BSc programmes
from the University of London International Foundation Programme
Students of the University of London International Foundation Programme (IFP) may progress to the BSc programmes by applying to transfer their registration, provided that they have passed all four full courses in the University of London International Foundation Programme.
Graduate Entry route
For graudate entry, in addition to the qualifications above, applicants must hold a full first degree completed in a minimum of three years duration on a full-time basis (or equivalent) from a university or other institution acceptable to the University of London.
Note: Applicants holding professional and/or postgraduate qualifications without a full first degree cannot be considered for admission to the Graduate Entry Route.
More information about the requirement:
List of Qualification for Entrance
List of Proficiency in English
For any enquiries or details, please contact 3761-1122 or email londonu@hkuspace.hku.hk.
Marketing Management (Module from Bachelor of Science in Economics and Management)
COURSE CODE 33Z13525A FEES $5,400 ENQUIRY 2867-8311
Information Systems and Organisations (Module from BSc Economics and Management)
COURSE CODE 23Z05248-4 FEES $5,400 ENQUIRY 2867-8311
Corporate Finance (Module from BSc Economics and Management)
Introduction to Economics (Module from Bachelor of Science in Economics and Management)
COURSE CODE 33Z106721 FEES $5,400 ENQUIRY 2867-8319
Principles of Accounting (Module from Bachelor of Science in Economics and Management)
Business and Management in a Global Context (Module from Bachelor of Science in Economics and Management)
Core Management Concepts (Module from Bachelor of Science in Economics and Management)
Microeconomics (Module from Bachelor of Science in Economics and Management)
Managerial Economics (Module from Bachelor of Science in Economics and Management)
Macroeconomics (Module from Bachelor of Science in Economics and Management)
Asset Pricing and Financial Markets (Module from Bachelor of Science in Economics and Management)
Monetary Economics (Module from Bachelor of Science in Economics and Management)
Strategy (Module from Bachelor of Science in Economics and Management)
Organisation Theory: an interdisciplinary approach (Module from Bachelor of Science in Economics and Management)
Human Resource Management (Module from Bachelor of Science in Economics and Management)
The Law of Business Organisations (Module from Bachelor of Science in Economics and Management)
Management and Innovation of E-business (Module from Bachelor of Science in Economics and Management)
Management Accounting (Module from Bachelor of Science in Economics and Management)
Principles of Corporate Finance (Module from Bachelor of Science in Economics and Management)
Principles of Banking and Finance (Module from Bachelor of Science in Economics and Management)
Financial Intermediation (Module from Bachelor of Science in Economics and Management)
Financial Reporting (Module from Bachelor of Science in Economics and Management)
Continuing Education Fund (Selected modules only)
For CEF courses that commence on or after 1 August 2022, 80% of the tuition fee for the first HK$10,000 subsidy and 60% of the tuition fee for the remaining HK$15,000 subsidy, subject to a maximum sum of HK$25,000 can be reimbursed on successful completion of an approved course.
* Students enrolled in the course are required to fulfill a minimum of 70% attendance and 50% overall mark of final examination.
Further information: http://www.wfsfaa.gov.hk/cef/
Continuing Education Fund Reimbursable Course (selected modules only)
Progression Path
Application Form Application Form
Enrolment Method
Please refer to the link below:
http://hkuspace.hku.hk/londonu#how-to-apply
University of London Business Programmes prospectus
University of London Business Programmes Application Deadline (for 2023/24 Semester):
Enrolment for HKU SPACE Tuition Course for 2023/24 Semester
Applicant is required to complete and return the Course Enrolment Form (printed in the centre of the brochure) with the course fee to any of our enrolment centres on a first-come first-served basis. The courses will start in early / mid September 2023.
Enrolment Deadline for HKU SPACE tuition courses for 2023/24 Semester:
Early September 2023 (First-come-first-served)
University of London fee - Application Fee & Registration Fee
Must be paid in pounds sterling, and may be paid either by a credit card recognized by MasterCard International or by the Visa group OR by banker's draft, cheque or UK postal order, made payable to the "University of London" and crossed 'a/c payee'. In addition, HKU SPACE also accepts cash, EPS, WeChat Pay, Alipay, VISA and Mastercard in HK$.
University of London Programmes (Admissions Office)
HKU SPACE
Room 313, 3/F, Admiralty Centre
18 Harcourt Road, Hong Kong
HKU SPACE Preparatory Course fee
Can be paid by cash / EPS / WeChat Pay / Alipay / VISA / Mastercard / Cheque in any of HKU SPACE enrolment centres.
Investment Management Business Management
BSc Accounting and Finance BSc Banking and Finance BSc Business and Management Advanced Diploma in Business Management
SHARING HKU SPACE Student beats 800 University of London candidates to gain global top spot
SHARING Ms Leung Kwan
SHARING Ms Lee Bong Siu Daisy
SHARING Ms Tsiang Shin Ting
BSc Accounting and Finance
BSc Banking and Finance
BSc Business and Management
Advanced Diploma in Business Management
Is the award conferred by the University of London?
What is the medium of instruction?
Can the students apply for any financial assistance and loans?
What is the assessment method?
Are the students required to comply with the HKU SPACE Vaccine Pass policy?

redpajama_set_name: RedPajamaCommonCrawl


BLUESIL MF 175 USP is a silicone Master batch, heat curing after addition of a vulcanization agent (chosen according to the processing method).
· Very slight replastification after prolonged storage.
· Very good compromise of mechanical properties and outstanding tensile strength in particular.
Extrusion, compression and injection moulding of translucent or colored articles requiring a good compromise of mechanical properties and a high tensile strength in particular.


text:
Home • Success Stories • Optimizely
Optimizing success for businesses across EMEA
Founded in 2010 and headquartered in San Francisco, Optimizely makes website optimization software for companies. In less than two years, the company's initial product has passed established players to become the leading A/B and multivariate testing platform in the world, and more than 6,000 enterprises have adopted Optimizely, including Starbucks, Disney and Salesforce.
"The number one reason we chose Amsterdam as our first location in Europe was its proximity to other markets," said Matt Althauser, General Manager of Optimizely EMEA. "With Schiphol, we're connected to our customers everywhere in EMEA, and there are daily flights back to our worldwide headquarters in San Francisco. That central location, and the multilingual nature of the people here, means that we can do business with anyone, anywhere." The presence of the international Internet exchange and a super-fast Internet connection was also a mission-critical reason to be in Amsterdam.
"When we were looking at where we wanted to locate in Europe, the Netherlands was the only country that had a service organization that helped us make that decision," Althauser added. "We worked with Amsterdam In Business and the NFIA, both in the US and in the Netherlands, and they engaged with us from the beginning. They gave us a tour of the Microsoft campus [near Schiphol], we also toured a server farm, and they even helped us scout potential office locations in Amsterdam."
Optimizely is growing quickly, and has big plans for its future in the Netherlands. Their Dutch presence has already grown to 20 people in just over a year, and they want to double that number in the coming year. "We have no trouble attracting new employees when we're based in a great city like Amsterdam!"
Location Amsterdam
Operations Headquarters, Marketing & Sales, Service Centers
Industry IT
"The number one reason we chose Amsterdam as our first location in Europe was its proximity to other markets. That central location, and the multilingual nature of the people here, means that we can do business with anyone, anywhere." Matt Althauser General Manager, Optimizely EMEA


text:
NewsPolitical
Michigan's redistricting maps raising concerns for black voters
The first set of drafts for Michigan's redistricting maps are already raising concern about how well they represent the African American community and whether they favor one party over the other.
By: Faraz Javed
(WXYZ) — The first set of drafts for Michigan's redistricting maps are already raising concern about how well they represent the African American community and whether they favor one party over the other.
Concerned legislators are urging Detroiters to step up before it's too late.
Democratic Michigan Senator Adam Hollier says the issue is not about red or blue state, it's about representing one of the largest black cities in the country.
"Black people make 14% of the state's population and 12% of the legislators, two-thirds of those members are from Detroit. If we decimate how black people get elected in this state it will have a far-reaching impact across the state," says Adam Hollier.
The same sentiment is being echoed by Republicans like former Majority Leader Michigan House Representatives Rocky Raczkowski.
"We are 100% behind the senator and his press conference because it's not about race, or color, or religion, it's about communities of interest and that the law is followed," says Raczkowski.
After being fed up with gerrymandering, voters took it upon themselves to create districts that better represent Michiganders of this decade.
In 2021, for the first time, the map-drawing process is being handled by a 13-member independent citizens commission instead of politicians.
The 10 maps include three House and Senate as well as four congressional district maps.
Next week, the maps will go on a state-wide tour where the public can ask questions and submit comments online.
The drafts will be voted on November 5th, followed by a 45-day public review period and a final vote on December 30th.
Also, the maps must receive a majority vote with bipartisan support, but even then Oakland University's Political Science Professor David Dulio says the process is far from over.
"Not only could there be revisions to the draft maps, but that's not the end of the process, someone is going to file a lawsuit," says Dulio.
Meanwhile, the public hearing for Detroiters will take place on October 20th at the TCF Center.


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also to deeds. Wherefore a gloss on Rm. 1:30, "contumelious, proud,"
something to the knowledge of others, they may do many kinds of harm.
unjustly against the respect due to the hearer.
honor due to him for any kind of excellence. This is done by "upbraiding"
(Ecclus. 20:15): "He will give a few things and upbraid much."
Nevertheless these terms are sometimes employed one for the other.
pertains to rail well, according to the Philosopher (Ethic. iv, 8).
Therefore railing or reviling is not a mortal sin.
or railing is not a mortal sin, by reason of its genus.
in fun, he would not be without blame.
word to give but little pain.
OBJ 1: It would seem that one ought not to suffer oneself to be reviled.
reviled, but rather reply to the reviler.
ought not to suffer another to be reviled, wherefore it is written (Prov.
neither should one suffer oneself to be reviled.
submit to reviling words, but rather answer back.
deaf man, heard not; and as a dumb man not opening his mouth."
prevented from progressing in virtue on account of our being reviled.
his folly, lest thou be like him."
not wood upon his fire."
reviling does not arise from anger.
as stated above (Q, A); whereas anger is opposed to meekness.
Therefore reviling does not arise from anger.
another is to revile him. Therefore reviling arises chiefly from anger.
excellency. Hence reviling does not arise directly from pride.
it an affront to themselves whenever anything is done against their will.
from folly on account of the latter's kinship with anger.