Nucleic Acids Research, Vol. 18, No. I

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Cloning of two distinct copies of human phosphoribosylpyrophosphate synthetase cDNA Blake J.Roessler, George Bell1, Steven Heidler1, Steven Seinol, Michael Becker1 and Thomas D.Palella Multipurpose Arthritis Center, Rackham Arthritis Research Unit and the Department of Internal Medicine, University of Michigan Medical School, Ann Arbor, Ml 48109 and 'University of Chicago and the Howard Hughes Medical Institute, Chicago, IL 60637, USA EMBL accession

Submitted October 4, 1989 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). GACTTCGGTTCCCCTCTCTCCAGCAGCCGTCATCCCTTACTCCAGYGCTTAG

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CCCCTCGCTCCtCCSCTGTCCTCCCCCACCTCCTCCCCCCCCCCGCGCCCCTCZGGAG I IF S G S H O D

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TATCCCAGCGCCTGCCCCACCGCCTGCCCCTCCACCTCCGCAACCTCCTTACCAACA TN V L SIO I A D L C L E L C F ACT

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ACCCCGCACTCCT0CACTCGATTCCGAAAACATTCCC 0CGAAAACTGTATCA IN a C I Z P A V L QN IE a

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MTAGAGTAACCTATTCATCACAAATTCACCACAACACCCCGCTTGCTCCACTGTAC

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MATCCAGAATCCCAAGTCTCCACCAACCCTACTCTCACTTCTCACTTCTTTTTCTT

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1. Palella,T.D. and Fox,I.H. (1989) In Scriver, C.S., et al. (eds.), The Metabolic Basis of Inherited Diseases. McGraw Hill Book Co., New York, 6th edition, pp. 965-1006. 2. Taira,M., Ishijima,S., Kita,K., Yamada,K., Iizasa,T. and Tatibana,M. (1987) J. Biol. Chem. 262, 14867-14872. 3. Iizasa,T., Taira,M., Shimada,H., Ishijima,S. and Tatibana,M. (1989) FEBS Lett., 244, 47-50. 4. Lee,C.C., Wu,X., Gibbs,R.A., Cook,R.G., Muzny,D.M. and Caskey,C.T. (1988) Science 239, 1288-1291. 5. Sanger,F., Nicklen,S. and Coulson,A.R. (1977) Proc. Natl. Acad. Sci. USA 74, 5463-5467.

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REFERENCES

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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).

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ACKNOWLEDGEMENTS

ATGTGGAATTTGCTTTGATCCACAAAGACACCAAGAACCCCAATGAAC TGACCGCA

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X15331, Y0097

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

?TCCGCCCCCCACCAtGCCCAACATCGCCTGTTCAGCCCCACCTCCCATCAGCACC N D P I V L F S G S S H I A D N L S L G L tATCTCACAAAATTGCTCACCGCC CCGCCT

nos

499

CTTTCTACATCCCACATCAGATATTAGACCTTATCCCAACTCCCAACACCCATT T?CTCCTTTTTACCTCTAGGTATTCAGCAATCATACGTTAATCACTGCCAAAAGCA GAGATTAACTCTO?CACCTCCTACCTGCATTATC?CATTCTGCCTTCCTTCATAATT TCACATCTTTCTATATOCTACATTTATTGTTTCCCTTCTAAACCTCAACATCATTT CTTTCAGTTTTCGAA

193