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. 1990 Aug;172(8):4555–4562. doi: 10.1128/jb.172.8.4555-4562.1990

Genes of the Escherichia coli pur regulon are negatively controlled by a repressor-operator interaction.

B He 1, A Shiau 1, K Y Choi 1, H Zalkin 1, J M Smith 1
PMCID: PMC213288  PMID: 2198266

Abstract

Fusions of lacZ were constructed to genes in each of the loci involved in de novo synthesis of IMP. The expression of each pur-lacZ fusion was determined in isogenic purR and purR+ strains. These measurements indicated 5- to 17-fold coregulation of genes purF, purHD, purC, purMN, purL, and purEK and thus confirm the existence of a pur regulon. Gene purB, which encodes an enzyme involved in synthesis of IMP and in the AMP branch of the pathway, was not regulated by purR. Each locus of the pur regulon contains a 16-base-pair conserved operator sequence that overlaps with the promoter. The purR product, purine repressor, was shown to bind specifically to each operator. Thus, binding of repressor to each operator of pur regulon genes negatively coregulates expression.

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Selected References

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  1. Aiba A., Mizobuchi K. Nucleotide sequence analysis of genes purH and purD involved in the de novo purine nucleotide biosynthesis of Escherichia coli. J Biol Chem. 1989 Dec 15;264(35):21239–21246. [PubMed] [Google Scholar]
  2. Bachmann B. J. Pedigrees of some mutant strains of Escherichia coli K-12. Bacteriol Rev. 1972 Dec;36(4):525–557. doi: 10.1128/br.36.4.525-557.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Benson C. E., Gots J. S. Occurrence of a regulatory deficiency in purine biosynthesis among pur A mutants of Salmonella typhimurium. Mol Gen Genet. 1976 Apr 23;145(1):31–36. doi: 10.1007/BF00331554. [DOI] [PubMed] [Google Scholar]
  4. Chan B., Busby S. Recognition of nucleotide sequences at the Escherichia coli galactose operon P1 promoter by RNA polymerase. Gene. 1989 Dec 14;84(2):227–236. doi: 10.1016/0378-1119(89)90496-4. [DOI] [PubMed] [Google Scholar]
  5. Choi K. Y., Zalkin H. Regulation of Escherichia coli pyrC by the purine regulon repressor protein. J Bacteriol. 1990 Jun;172(6):3201–3207. doi: 10.1128/jb.172.6.3201-3207.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Ebbole D. J., Zalkin H. Cloning and characterization of a 12-gene cluster from Bacillus subtilis encoding nine enzymes for de novo purine nucleotide synthesis. J Biol Chem. 1987 Jun 15;262(17):8274–8287. [PubMed] [Google Scholar]
  7. Fath M. J., Mahanty H. K., Kolter R. Characterization of a purF operon mutation which affects colicin V production. J Bacteriol. 1989 Jun;171(6):3158–3161. doi: 10.1128/jb.171.6.3158-3161.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Flannigan K. A., Hennigan S. H., Vogelbacker H. H., Gots J. S., Smith J. M. Purine biosynthesis in Escherichia coli K12: structure and DNA sequence studies of the purHD locus. Mol Microbiol. 1990 Mar;4(3):381–392. doi: 10.1111/j.1365-2958.1990.tb00605.x. [DOI] [PubMed] [Google Scholar]
  9. Gots J. S., Benson C. E., Jochimsen B., Koduri K. R. Microbial models and regulatory elements in the control of purine metabolism. Ciba Found Symp. 1977;(48):23–41. doi: 10.1002/9780470720301.ch3. [DOI] [PubMed] [Google Scholar]
  10. Hawley D. K., McClure W. R. Compilation and analysis of Escherichia coli promoter DNA sequences. Nucleic Acids Res. 1983 Apr 25;11(8):2237–2255. doi: 10.1093/nar/11.8.2237. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Houlberg U., Jensen K. F. Role of hypoxanthine and guanine in regulation of Salmonella typhimurium pur gene expression. J Bacteriol. 1983 Feb;153(2):837–845. doi: 10.1128/jb.153.2.837-845.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Keilty S., Rosenberg M. Constitutive function of a positively regulated promoter reveals new sequences essential for activity. J Biol Chem. 1987 May 5;262(13):6389–6395. [PubMed] [Google Scholar]
  13. Kilstrup M., Meng L. M., Neuhard J., Nygaard P. Genetic evidence for a repressor of synthesis of cytosine deaminase and purine biosynthesis enzymes in Escherichia coli. J Bacteriol. 1989 Apr;171(4):2124–2127. doi: 10.1128/jb.171.4.2124-2127.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kohara Y., Akiyama K., Isono K. The physical map of the whole E. coli chromosome: application of a new strategy for rapid analysis and sorting of a large genomic library. Cell. 1987 Jul 31;50(3):495–508. doi: 10.1016/0092-8674(87)90503-4. [DOI] [PubMed] [Google Scholar]
  15. Levine R. A., Taylor M. W. Mechanism of adenine toxicity in Escherichia coli. J Bacteriol. 1982 Mar;149(3):923–930. doi: 10.1128/jb.149.3.923-930.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Levine R. A., Taylor M. W. Regulation of purE transcription in a purE::lac fusion strain of Escherichia coli. J Bacteriol. 1982 Mar;149(3):1041–1049. doi: 10.1128/jb.149.3.1041-1049.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Levine R. A., Taylor M. W. Selection for purine regulatory mutants in an E. coli hypoxanthine phosphoribosyl transferase-guanine phosphoribosyl transferase double mutant. Mol Gen Genet. 1981;181(3):313–318. doi: 10.1007/BF00425604. [DOI] [PubMed] [Google Scholar]
  18. Makaroff C. A., Zalkin H. Regulation of Escherichia coli purF. Analysis of the control region of a pur regulon gene. J Biol Chem. 1985 Aug 25;260(18):10378–10387. [PubMed] [Google Scholar]
  19. Meng L. M., Kilstrup M., Nygaard P. Autoregulation of PurR repressor synthesis and involvement of purR in the regulation of purB, purC, purL, purMN and guaBA expression in Escherichia coli. Eur J Biochem. 1990 Jan 26;187(2):373–379. doi: 10.1111/j.1432-1033.1990.tb15314.x. [DOI] [PubMed] [Google Scholar]
  20. Nonet M. L., Marvel C. C., Tolan D. R. The hisT-purF region of the Escherichia coli K-12 chromosome. Identification of additional genes of the hisT and purF operons. J Biol Chem. 1987 Sep 5;262(25):12209–12217. [PubMed] [Google Scholar]
  21. Ponnambalam S., Chan B., Busby S. Functional analysis of different sequence elements in the Escherichia coli galactose operon P2 promoter. Mol Microbiol. 1988 Mar;2(2):165–172. doi: 10.1111/j.1365-2958.1988.tb00018.x. [DOI] [PubMed] [Google Scholar]
  22. Rolfes R. J., Zalkin H. Escherichia coli gene purR encoding a repressor protein for purine nucleotide synthesis. Cloning, nucleotide sequence, and interaction with the purF operator. J Biol Chem. 1988 Dec 25;263(36):19653–19661. [PubMed] [Google Scholar]
  23. Rolfes R. J., Zalkin H. Regulation of Escherichia coli purF. Mutations that define the promoter, operator, and purine repressor gene. J Biol Chem. 1988 Dec 25;263(36):19649–19652. [PubMed] [Google Scholar]
  24. Schendel F. J., Mueller E., Stubbe J., Shiau A., Smith J. M. Formylglycinamide ribonucleotide synthetase from Escherichia coli: cloning, sequencing, overproduction, isolation, and characterization. Biochemistry. 1989 Mar 21;28(6):2459–2471. doi: 10.1021/bi00432a017. [DOI] [PubMed] [Google Scholar]
  25. Shiau A., Smith J. M. Improved cat gene cassette for promoter analysis and genetic constructions. Gene. 1988 Jul 30;67(2):295–299. doi: 10.1016/0378-1119(88)90406-4. [DOI] [PubMed] [Google Scholar]
  26. Smith J. M., Daum H. A., 3rd Nucleotide sequence of the purM gene encoding 5'-phosphoribosyl-5-aminoimidazole synthetase of Escherichia coli K12. J Biol Chem. 1986 Aug 15;261(23):10632–10636. [PubMed] [Google Scholar]
  27. Thomulka K. W., Gots J. S. Isolation and characterization of purine regulatory mutants of Salmonella typhimurium with an episomal purE-lac fusion. J Bacteriol. 1982 Jul;151(1):153–161. doi: 10.1128/jb.151.1.153-161.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Tiedeman A. A., Keyhani J., Kamholz J., Daum H. A., 3rd, Gots J. S., Smith J. M. Nucleotide sequence analysis of the purEK operon encoding 5'-phosphoribosyl-5-aminoimidazole carboxylase of Escherichia coli K-12. J Bacteriol. 1989 Jan;171(1):205–212. doi: 10.1128/jb.171.1.205-212.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Tiedeman A. A., Smith J. M. lacZY gene fusion cassettes with KanR resistance. Nucleic Acids Res. 1988 Apr 25;16(8):3587–3587. doi: 10.1093/nar/16.8.3587. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Tso J. Y., Zalkin H., van Cleemput M., Yanofsky C., Smith J. M. Nucleotide sequence of Escherichia coli purF and deduced amino acid sequence of glutamine phosphoribosylpyrophosphate amidotransferase. J Biol Chem. 1982 Apr 10;257(7):3525–3531. [PubMed] [Google Scholar]
  31. VOGEL H. J., BONNER D. M. Acetylornithinase of Escherichia coli: partial purification and some properties. J Biol Chem. 1956 Jan;218(1):97–106. [PubMed] [Google Scholar]
  32. Vidal-Ingigliardi D., Raibaud O. A convenient technique to compare the efficiency of promoters in Escherichia coli. Nucleic Acids Res. 1985 Aug 26;13(16):5919–5926. doi: 10.1093/nar/13.16.5919. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Vieira J., Messing J. Production of single-stranded plasmid DNA. Methods Enzymol. 1987;153:3–11. doi: 10.1016/0076-6879(87)53044-0. [DOI] [PubMed] [Google Scholar]
  34. Watanabe W., Sampei G., Aiba A., Mizobuchi K. Identification and sequence analysis of Escherichia coli purE and purK genes encoding 5'-phosphoribosyl-5-amino-4-imidazole carboxylase for de novo purine biosynthesis. J Bacteriol. 1989 Jan;171(1):198–204. doi: 10.1128/jb.171.1.198-204.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Way J. C., Davis M. A., Morisato D., Roberts D. E., Kleckner N. New Tn10 derivatives for transposon mutagenesis and for construction of lacZ operon fusions by transposition. Gene. 1984 Dec;32(3):369–379. doi: 10.1016/0378-1119(84)90012-x. [DOI] [PubMed] [Google Scholar]
  36. Wilson H. R., Chan P. T., Turnbough C. L., Jr Nucleotide sequence and expression of the pyrC gene of Escherichia coli K-12. J Bacteriol. 1987 Jul;169(7):3051–3058. doi: 10.1128/jb.169.7.3051-3058.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Wilson H. R., Turnbough C. L., Jr Role of the purine repressor in the regulation of pyrimidine gene expression in Escherichia coli K-12. J Bacteriol. 1990 Jun;172(6):3208–3213. doi: 10.1128/jb.172.6.3208-3213.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Winans S. C., Elledge S. J., Krueger J. H., Walker G. C. Site-directed insertion and deletion mutagenesis with cloned fragments in Escherichia coli. J Bacteriol. 1985 Mar;161(3):1219–1221. doi: 10.1128/jb.161.3.1219-1221.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Wolfe S. A., Smith J. M. Separate regulation of purA and purB loci of Escherichia coli K-12. J Bacteriol. 1985 May;162(2):822–825. doi: 10.1128/jb.162.2.822-825.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]

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