1887

Abstract

The gene, encoding phosphoribosylpyrophosphate synthetase, is preceded by a leader, which is 302 bp long in and 417 bp in . A potential open reading frame (ORF) extends across the promoter and into the leader. The region between the coding region and an upstream gene () in and was cloned, sequenced and shown to encode two ORFs of unknown function. ORF 1 encodes a 23 kDa protein and ORF 2 a 31 kDa protein, as observed by denaturing PAGE of extracts of cells bearing plasmids encoding the ORFs. Both ORFs are transcribed in the same direction as the gene with ORF 2 extending into the leader. Northern blot analysis showed that the message in was on 1·3 and 2·7 kb transcripts. The shorter transcript encoded the gene only, while the longer transcript also encoded the two ORFs. Thus, the gene is transcribed from two promoters, the first promoter (P) originating upstream of ORF 1, and expressing the gene in a tricistronic operon and a second promoter (P), located within the ORF 2 coding frame, which transcribes the gene only. The transcripts encoding only were 20 times as abundant as the tricistronic transcripts under all conditions examined. This was the case whether cells containing plasmid-encoded or only chromosomally encoded copies of the region were probed for these transcripts. Derepression of the gene upon pyrimidine starvation was shown to be due to an increase in the amount of message originating from the promoter P.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-139-2-259
1993-02-01
2021-07-30
Loading full text...

Full text loading...

/deliver/fulltext/micro/139/2/mic-139-2-259.html?itemId=/content/journal/micro/10.1099/00221287-139-2-259&mimeType=html&fmt=ahah

References

  1. Bower S.G., Hove-Jensen B., Switzer R.L. 1988; Structure of the gene encoding phosphoribosylpyrophosphate synthetase (prsA) in Salmonella typhimurium.. Journal of Bacteriology 170:3243–3248
    [Google Scholar]
  2. Bower S.G., Harlow K.W., Switzer R.L., Hove-Jensen B. 1989; Characterization of the Escherichia coli prsAl-encoded mutant phosphoribosyl pyrophosphate synthetase identifies a divalent cation-nucleotide binding site. Journal of Biological Chemistry 264:10287–10291
    [Google Scholar]
  3. Clark D.J., Maaløe O. 1967; DNA replication and the division cycle in Escherichia coli. Journal of Molecular Biology 23:99–112
    [Google Scholar]
  4. Davis R.W., Botstein D., Roth J.R. 1980 Advanced Bacterial Genetics Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  5. Drolet M., Peloquin L., Echelard Y., Cousineau L., Sasarman A. 1989; Isolation and nucleotide sequence of the hem A gene of Escherichia coliK 12. Molecular and General Genetics 216:347–352
    [Google Scholar]
  6. Elliott T. 1989; Cloning, genetic characterization, and nucleotide sequence of the hemA-prfA operon of Salmonella typhimurium. Journal of Bacteriology 171:3948–3960
    [Google Scholar]
  7. Hove-Jensen B. 1985; Cloning and characterization of the prs gene encoding phosphoribosylpyrophosphate synthetase ofEscherichia coli. Molecular and General Genetics 201:269–276
    [Google Scholar]
  8. Hove-Jensen B., Harlow K.H., King C.J., Switzer R.L. 1986; Phosphoribosylpyrophosphate synthetase of Escherichia coli : Properties of the purified enzyme and primary structure of the prs gene. Journal of Biological Chemistry 261:6765–6771
    [Google Scholar]
  9. Laemmli U.K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature; London: 227209–217
    [Google Scholar]
  10. Li J.-M., Russell C.S., Cosloy S.D. 1989; Cloning and structure of the hem A gene of Escherichia coli K-12. Gene 82209–217
    [Google Scholar]
  11. Matsudaira P. 1987; Sequence from picomole quantities of proteins electroblotted onto polyvinylidene difluoride membranes. Journal of Biological Chemistry 262:10035–10038
    [Google Scholar]
  12. Miller J.H. 1972 Experiments in Molecular Genetics Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  13. Misuzawa S., Ward D.F. 1982; A bacteriophage lambda vector for cloning with BamELl and Sau3A . Gene 20:317–322
    [Google Scholar]
  14. Neuhard J., Nygaard P. 1987; Purines and pyrimidines. In Escherichia coli and Salmonella typhimurium Cellular and Molecular Biology 1 pp. 445–473 Edited by Ingraham J.L., Low K. B., Neidhardt F.C., Magasanik B., Schaechter M., Umbarger H.E. Washington DC: American Society for Microbiology.;
    [Google Scholar]
  15. Nilsson D., Hove-Jensen B. 1987; Phosphoribosylpyrophosphate synthetase ofBacillus subtilis. Cloning, characterization and chromosomal mapping of the prs gene. Gene 53:247–255
    [Google Scholar]
  16. Olszowy J., Switzer R.L. 1972; Specific repression of phosphoribosylpyrophosphate synthetase by uridine compounds in Salmonella typhimurium. . Journal of Bacteriology 110:450–451
    [Google Scholar]
  17. Post D., Switzer R.L. 1991; prsB is an allele of the Salmonella typhimurium prsA gene:characterization of a mutant phosphoribosylpyrophosphate synthetase. Journal of Bacteriology 173:1978–1986
    [Google Scholar]
  18. Promega Corporation 1991 Catalogue and Reference Guide- Biological Research Products pp. 58–61 Madison: WI;
    [Google Scholar]
  19. Sambrook J., Fritsch E.F., Maniatis T. 1989 Molecular Cloning: a Laboratory Manual, 2nd edn.. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  20. Sanger F., Coulsen A.R., Barrell B.G., Smith A.J.H., Roe B.A. 1980; Clonging in single-stranded bacteriophage as an aid to rapid DNA sequencing. Journal of Molecular Biology 143:161–178
    [Google Scholar]
  21. Schleif R.F., Wensink P.C. 1981 Practical Methods in Molecular Biology New York: Springer-Verlag;
    [Google Scholar]
  22. Silhavy T.J., Berman M.L., Enquist L.W. 1984 Experiments with Gene Fusions Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  23. Sorenson M.A., Kurland C.G., Pedersen S. 1989; Codon usage determines translation rate in Escherichia coli. Journal of Molecular Biology 207:365–377
    [Google Scholar]
  24. Sternberg N., Tiemeier D., Enquist L. 1977; In vitro packaging of a lambdaDam vector containing EcoRI DNA fragments of Escherichia coli and phage PI. Gene 1:255–280
    [Google Scholar]
  25. Verkamp E., Chelm B.K. 1989; Isolation, nucleotide sequence, and preliminary characterization of theEscherichia coli K-12hem Agene. Journal of Bacteriology 171:4728–4735
    [Google Scholar]
  26. White M.N., Olszowy J., Switzer R.L. 1971; Regulation and mechanism of phosphoribosylpyrophosphate synthetase: repression by end products. Journal of Bacteriology 108:122–131
    [Google Scholar]
  27. Yanisch-Perron C., Vieira J., Messing J. 1985; Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mpl8 and pUC19 vectors. Gene 33:103–119
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-139-2-259
Loading
/content/journal/micro/10.1099/00221287-139-2-259
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error