Cloning of two distinct copies of human phosphoribosylpyrophosphate synthetase cDNA


Phosphoribosylpyrophosphate synthetase (PRS) catalyzes the phosphoribosylation of ribose 5-phosphate to 5-phosphoribosyl-1-pyrophosphate (PRPP), which is necessary for the de novo and salvage pathways of purine, pyrimidine and pyridine biosynthesis (1). In the rat, two nearly identical cDNA transcripts (PRS I and PRS II) have been cloned (2). Additionally, a human cDNA homologous to the rat PRS cDNA has been cloned (3). We report the isolation of a human PRS cDNA homologous to rat PRS I and confirm the nucleotide sequence of an independently isolated human PRS cDNA. Published rat cDNA sequences were used to design flanking oligonucleotide primers, and a partial length human PRS cDNA probe was generated from human lymphoblast mRNA by the polymerase chain reaction (4). Human PRS I was isolated from lymphoblast cDNA libraries cloned into lambda gtlO. Human PRS II was isolated from human kidney cell cDNA libraries cloned into lambda gtlO. Both human PRS I and PRS II cDNA were sequenced by the Sanger method (5). 
TATCCCAGCGCCTGCCCCACCGCCTGCCCCTCCACCTCCGCAACCTCCTTACCAACA L S IO I A D L C L E L C V TN F S N O E T C V C S V CE D ACT TCACAACCACCACACCTCTCTCCAAATCGCTCTCCTACCTGCACACAATC ACTTCACCAACCACCACACCACCCTGCAGATTCCTCAAACCCTCACAGGCCAAGATC F S T C V I C * V a C D V Y I VOS C C G I L N L L
TCTACATTCTTCACACTCCTTCTGCCGSAAACAAT.ACAATTTAATCCAGCTTTTGA ATGTGGAATTTGCTTTGATCCACAAAGACACCAAGAACCCCAATGAACTGACCGCA 
P P D AG GA K K R V T S I A D R L K V D F A L I N K E K K K A K E V D
K V E F A L I N K C R K K A K E V D K YV L V G D V K D K V A I L V D



  L G L E L G K V V TtATCTCACAAAATTGCTCACCGCC CCGCCT CAGCTACCCAACGCGTGTACTAACA :~!t..... ....... *! ... ...........*--*-..-*** ... -10 




  ... . ..... o. .....** ...* .-. * .. .... ..-.. -*. s****9 ** . -*-...-------*-.. .. --..........-........ .. ..... -.*. *-*-***-^* -...................... ............. 556 TTGTTTCACCTGACGCACCCGGACCAACACCCCTTACATCAATTCCAGACAGGTTGA I V 




  R ATCTGGATTTGCCtTTGATTCACAAAGACGCCAA.CAACCCAATGAAGTGGACCCCA . . . . . . . . . . . ........ . ... . --.-*--* *** .... ......................-. 613 


4 .

 4 . .. ..... ........T.....T..C..........A ... ...... z V G D V K D R V A I L V D D N A D T C C T I C N A A D K L L S A C A T ACACTTGTGCCACAATCTCCCATCCAGCTCACAAACTTCTCTCAGCTGCCCCCACCA .t. .. . .. .. ... *..* .-.*** -* .... .. .. ........ ....... A I L T N C I F S C P A I S R I N A C F E A V VV T N T I t Q t D K ACAAC CATCCTTTGACCCACTACTAGTCACCAATACCATACCTCACGAGCACAAGA *.! ... *. .. . .. ** .. * ... .. . ... ... . 41 ATAATGCCCCCTTTGAGGCTGTTGTCGTCACAAACACAATTCCGCAAGACGACAAAA K N A AF E A V V V T N T I P O E D K NK N C S K I O V I D IS N I L A E A TCGACCATTCCTCCAAAATACACGTCATTCACATCTCTATCATCCTTCCACAAGCCA . . . . . ...... ....... .......... ...... ..... ........ .. .. ... ..... . 999 ?CAAACACTCCACCAAGATTCACCTCATTCACATTTCCATCATCTTCCCCAACAA N N C S K I Q V I D I S N I L A E A It AN TN N C E S V S Y L F S H V P L TCACCACAACTCACATGGACAATCCCTTTCTTACCTATTCAGCCATCTCCCTTTAZ *. . .. .. ........ ........ e. ..... .............. .. 9SS TCCCAACCACACACAATCCCCAATCCCTCTCCTACCTCTTCACCCATCTCCCCCTAI I T R N N K E S V S Y L F S N V P L MTAGAGTAACCTATTCATCACAAATTCACCACAACACCCCGCTTGCTCCACTGTAC . . . .-****.................1012 MATCCAGAATCCCAAGTCTCCACCAACCCTACTCTCACTTCTCACTTCTTTTTCTT CTTTCTACATCCCACATCAGATATTAGACCTTATCCCAACTCCCAACACCCATT T?CTCCTTTTTACCTCTAGGTATTCAGCAATCATACGTTAATCACTGCCAAAAGCA GAGATTAACTCTO?CACCTCCTACCTGCATTATC?CATTCTGCCTTCCTTCATAATT TCACATCTTTCTATATOCTACATTTATTGTTTCCCTTCTAAACCTCAACATCATTT CTTTCAGTTTTCGAA 


ACKNOWLEDGEMENTS

This research was supported by grants P60 AR20557 and R29 DK38932-01 from the NIH, NIAMS. These sequence data appear in the EMBL/GenBank/DDBJ Nucleotide Sequence Databases under the accession numbers X15331 (HPR SI) and Y00971 (HPR SIH).
Blake J Roessler, George Bell1, Steven Heidler1, Steven Seinol, Michael Becker1, Thomas D Palella
Cloning of two distinct copies of human phosphoribosylpyrophosphate synthetase cDNA