Skip to main content
Log in

Sequence analysis of the DdPYR5-6gene coding for UMP synthase in Dictyostelium discoideum and comparison with orotate phosphoribosyl transferases and OMP decarboxylases

  • Published:
Molecular and General Genetics MGG Aims and scope Submit manuscript

Summary

A Dictyostelium discoideum DNA fragment that complements the ura3 and the ura5 mutants of Saccharomyces cerevisiae has been sequenced. It contains an open reading frame of 478 codons capable of encoding a polypeptide of molecular weight 52475. This gene, named DdPYR5-6, encodes a bifunctional protein composed of the orotate phosphoribosyl transferase (OPRTase) and the orotidine-5′-phosphate decarboxylase (OMPdecase) domains described for UMP synthase in mammals. The existence of separate domains for the two activities was suspected because deletion of the N-terminal coding segment of the gene eliminated the ura5 but not the ura3 complementing activity. We have now confirmed that the two parts of the open reading frame share homology with known OPRTase and OMPdecase sequences. Several blocks of sequence are conserved among OPRTase from bacteria, fungi and slime mold and one of them corresponds to the consensus sequence for phosphoribosylbinding sites. The OMPdecase domain shows extensive similarity with the yeast and Neurospora crassa enzymes, suggesting that they have evolved from an ancestral gene which was fused to the OPRTase gene in D. discoideum. It is less related to the bacterial enzyme but all these sequences present conserved blocks of homology which could identify the active site. The codon usage is strongly biased in a manner similar to that found for other D. discoideum genes. The flanking DNA contains homopolymers of A and T and alternating sequences that are characteristic of the gene organization in D. discoideum.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Beckwith JR, Pardee AB, Austrian R, Jacob F (1962) Coordination of the synthesis of the enzymes in the pyrimidine pathway of E. coli. J Mol Biol 5:618–634

    Google Scholar 

  • Boy-Marcotte E, Jacquet M (1982) A Dictyostelium discoideum DNA fragment complements a Saccharomyces cerevisiae ura3 mutant. Gene 20:433–440

    Google Scholar 

  • Boy-Marcotte E, Vilaine F, Camonis J, Jacquet M (1984) A DNA sequence from Dictyostelium discoideum complements ura3 and ura5 mutations of Saccharomyces cerevisiae. Mol Gen Genet 193:406–413

    Google Scholar 

  • Brown GK, O'Sullivan WJ (1977) Subunit structure of the orotate phosphoribosyltransferase-orotidylate-decarboxylase complex. Biochemistry 16:321–332

    Google Scholar 

  • Caroline DF (1969) Pyrimidine biosynthesis in Neurospora crassa: gene enzyme relationships. J Bacteriol 100:1371–1377

    Google Scholar 

  • Chothia C, Lesk AM (1986) The use of sequence homologies to predict protein structures. In: Fletterick R, Zoller M (eds) Computer graphic and molecular modeling. Cold Spring Harbor Laboratory Press, New York, pp 33–37

    Google Scholar 

  • Faure M, Kalekine M, Boy-Marcotte E, Jacquet M (1987) Developmental control of the expression of the dihydroorotate dehydrogenase and UMP synthase genes in Dictyostelium discoideum. Cell Differ, in press

  • Hershey HV, Taylor MW (1986) Nucleotide sequence and deduced amino-acid sequence of E. coli adenine phosphoribosyl transferase and comparison with other analogous enzymes. Gene 43:287–293

    Google Scholar 

  • Jacquet M, Boy-Marcotte E, Rossier C, Kessin RH (1982) A fragment of Dictyostelium discoideum genomic DNA that complements the ura1 mutation of Saccharomyces cerevisiae. J Mol Appl Genet 1:513–525

    Google Scholar 

  • Jacquet M, Kalekine M, Boy-Marcotte E (1985) Sequence analysis of a Dictyostelium discoideum gene coding for an active dihydroorotate dehydrogenase in yeast. Biochimie 67:583–588

    Google Scholar 

  • Jones ME (1980) Pyrimidine nucleotide biosynthesis in animals: genes, enzymes and regulation of UMP biosynthesis. Annu Rev Biochem 49:253–279

    Google Scholar 

  • Kimmel AR, Firtel RA (1983) Sequence organisation in Dictyostelium: unique structure at the 5′-end of coding genes. Nucleic Acids Res 11:541–552

    Google Scholar 

  • Lacroute F (1968) Regulation of pyrimidine biosynthesis in S. cerevisiae. J Bacteriol 95:824–832

    Google Scholar 

  • Lilley D (1986) Bent molecules — How and why? Nature 320:487

    Google Scholar 

  • McCarroll R, Olsen GJ, Stahl YD, Woese CR, Sogin ML (1983) Nucleotide sequence of the Dictyostelium discoideum small-subunit ribosomal ribonucleic acid inferred from the gene sequence: evolutionary implications. Biochemistry 22:5858–5868

    Google Scholar 

  • Messing J, Crea R, Seeburg PH (1981) A system for shotgun DNA sequencing. Nucleic Acids Res 9:309–321

    Google Scholar 

  • Neuhard J, Stauning E, Kelln RA (1985) Cloning and characterization of the pyrE gene and the pyrE:: Mud1 (Aplac) fusions from Salmonella typhimurium. Eur J Biochem 146:597–603

    Google Scholar 

  • Newbury SF, Glazebrook JA, Radford A (1986) Sequence analysis of the pyr-4 (orotidine 5′-P decarboxylase) gene of Neurospora crassa. Gene 43:51–58

    Google Scholar 

  • Poulsen P, Jensen KF, Valentin-Hansen P, Carlsson P, Lundberg LG (1983) Nucleotide sequence of the Escherichia coli PyrE gene and of the DNA in front of the protein coding region. Eur J Biochem 135:223–229

    Google Scholar 

  • Proudfoot NJ, Brownlee GG (1976) 3′ non coding region sequences in eucaryotic messenger RNA. Nature 263:211–214

    Google Scholar 

  • Rawls JM (1978) The enzymes for de novo pyrimidine biosynthesis in Drosophila melanogaster: their localization, properties and expression during development. Biochem Physiol 628:207–216

    Google Scholar 

  • Rich A, Nordheim A, Wang AHJ (1984) The chemistry and biology of left-handed Z-DNA. Annu Rev Biochem 53:791–846

    Google Scholar 

  • Rose M, Grisafi P, Botstein D (1984) Structure and function of the yeast URA3 gene: expression in Escherichia coli. Gene 29:113–124

    Google Scholar 

  • Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467

    Google Scholar 

  • Schartz RM, Dayhoff MO (1979) Matrices for detecting distant relationships. In: Dayhoff MO (ed) Atlas of protein sequence and structure, vol 5 sup 3. National Biomedical Research Foundation, Washington D.C., pp 353–358

    Google Scholar 

  • Schwartz M, Neuhard J (1975) Control of expression of the pyr genes in S. thyphimurium: effect of variations in uridine and cytidien nucleotide pools. J Bacteriol 121:814–822

    Google Scholar 

  • Theisen M, Kelln RA, Neuahrd J (1987) Cloning and characterization of the pyrF operon of Salmonella typhimurium. Eur J Biochem 164:613–619

    Google Scholar 

  • Traut TW (1982) UMP synthase: the importance of quaternary structure in channeling intermediates. TIBS: 255–257

  • Turcq B, Bégueret J (1987) The ura5 gene of the filamentous fungus Podospora anserina: nucleotide sequence and expression in transformed strains. Gene 53:201–209

    Google Scholar 

  • Warrick HM (1987) Codon frequency in Dictyostelium discoideum. Methods Cell Biol 28:497–500

    Google Scholar 

  • Williams JG, Pears CJ, Jermyn KA, Driscoll DM, Mahbubani J, Kay R (1986) The control of gene expression during cellular differentiation of Dictyostelium discoideum. In: Both IR, Higgins CF (eds) The society for general microbiology, Symposium 39; Regulation of gene expression 25 years on Cambridge University Press, Cambridge pp 277–298

Download references

Author information

Authors and Affiliations

Authors

Additional information

Communicated by P. Tiollais

Rights and permissions

Reprints and permissions

About this article

Cite this article

Jacquet, M., Guilbaud, R. & Garreau, H. Sequence analysis of the DdPYR5-6gene coding for UMP synthase in Dictyostelium discoideum and comparison with orotate phosphoribosyl transferases and OMP decarboxylases. Molec. Gen. Genet. 211, 441–445 (1988). https://doi.org/10.1007/BF00425698

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00425698

Key words

Navigation