We observed an intriguing role of Cas5 on governing RNA PolII binding at genes important for cell cycle, meiosis, and DNA replication (Figs. 1c and 2c), suggesting Cas5 regulates cell cycle dynamics under basal and cell wall stress conditions. If this were the case, the expression pattern of cell cycle regulated genes would be perturbed in the absence of Cas5. Using the Candida cell cycle database33, we determined that 56 out of the 604 genes that had Cas5-dependent RNA PolII binding under basal conditions, were genes whose expression peaked during the cell cycle (Supplementary Data 6). Of these, over 75% (43 out of 56) had reduced RNA PolII binding in the cas5Δ/cas5Δ mutant as compared with the wild-type strain (Fig. 4a and Supplementary Data 6). Next, we analyzed the class of genes involved in cell cycle that had reduced RNA PolII binding upon caspofungin treatment (Fig. 1e). Among these genes were all six factors of the mini-chromosome maintenance (MCM) complex (Fig. 4b), which has important roles in the formation of the pre-replicative complex (pre-RC) prior to DNA replication34. Reduced RNA PolII binding at genes encoding MCM complex components was dependent on Cas5 (Fig. 4b). As expected, qRT-PCR confirmed that reduced expression of representative genes, MCM2 and MCM3, was also dependent on Cas5 (Fig. 4c). Finally, we classified genes that were differentially bound by RNA PolII in response to caspofungin based on their peak expression throughout the cell cycle, once again using the Candida cell cycle database33. In the wild-type strain, genes with increased RNA PolII binding in response to caspofungin were predominantly G1-specific, and those with reduced RNA PolII binding were predominantly M phase-specific (Fig. 4d and Supplementary Data 6). Importantly, these trends were less apparent in a mutant lacking Cas5 (Fig. 4d and Supplementary Data 6). Overall, our results highlight the important role of Cas5 in ensuring appropriate cell cycle dynamics under basal conditions and in response to cell wall stress.Fig. 4Cas5 governs cell cycle dynamics under basal and cell wall stress conditions. a Circos plot depicting the number of genes with increased (red) and decreased (blue) RNA PolII binding in the cas5∆/cas5∆ mutant whose normal expression in the wild-type peaks at the indicated cell cycle stage. b Cas5 is required for the decreased PolII binding at genes encoding MCM complex members and CDC6 in response to caspofungin. Histogram showing the PolII ChIP-Seq signal for the indicated genes encoding MCM complex components in response to caspofungin treatment. Error bars represent standard deviation (s.d.) from the mean of duplicate samples. c Downregulation of MCM2 and MCM3 in response to caspofungin depends on Cas5. Transcript levels were monitored by qRT-PCR and normalized to TEF1. Error bars represent s.d. from the mean of triplicate samples. Significance was measured with an unpaired t test in GraphPad Prism (*P < 0.05). d Circos plots illustrating the number of genes with increased (red) or decreased (blue) PolII binding in response to caspofungin (CF) in wild-type and cas5∆/cas5∆ strains for genes whose expression is regulated throughout the cell cycle