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. 1990 Oct;172(10):5758–5766. doi: 10.1128/jb.172.10.5758-5766.1990

Autoregulation of Escherichia coli purR requires two control sites downstream of the promoter.

R J Rolfes 1, H Zalkin 1
PMCID: PMC526892  PMID: 2211510

Abstract

The expression of Escherichia coli purR, which encodes the pur regulon repressor protein, is autoregulated. Autoregulation at the level of transcription requires two operator sites, designated purRo1 and purRo2 (O1 and O2). Operator O1 is in the region of DNA between the transcription start site and the site for translation initiation, and O2 is in the protein-coding region. The repressor protein binds noncooperatively to O1 with a sixfold-higher affinity than to O2, and saturation of O1 by the repressor precedes saturation of O2. Both O1 and O2 function in the two- to threefold autoregulation in vivo, as determined by measurement of beta-galactosidase and mRNA from purR-lacZ translational fusions. Of all the genes thus far known to be regulated by the Pur repressor, only purR employs a two-operator mechanism.

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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. Linkage map of Escherichia coli K-12, edition 7. Microbiol Rev. 1983 Jun;47(2):180–230. doi: 10.1128/mr.47.2.180-230.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bogosian G., Bertrand K., Somerville R. Trp repressor protein controls its own structural gene. J Mol Biol. 1981 Jul 15;149(4):821–825. doi: 10.1016/0022-2836(81)90361-2. [DOI] [PubMed] [Google Scholar]
  4. Casadaban M. J. Transposition and fusion of the lac genes to selected promoters in Escherichia coli using bacteriophage lambda and Mu. J Mol Biol. 1976 Jul 5;104(3):541–555. doi: 10.1016/0022-2836(76)90119-4. [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. Churchward G., Belin D., Nagamine Y. A pSC101-derived plasmid which shows no sequence homology to other commonly used cloning vectors. Gene. 1984 Nov;31(1-3):165–171. doi: 10.1016/0378-1119(84)90207-5. [DOI] [PubMed] [Google Scholar]
  7. Dandanell G., Valentin-Hansen P., Larsen J. E., Hammer K. Long-range cooperativity between gene regulatory sequences in a prokaryote. 1987 Feb 26-Mar 4Nature. 325(6107):823–826. doi: 10.1038/325823a0. [DOI] [PubMed] [Google Scholar]
  8. DiLauro R., Taniguchi T., Musso R., de Crombrugghe B. Unusual location and function of the operator in the Escherichia coli galactose operon. Nature. 1979 Jun 7;279(5713):494–500. doi: 10.1038/279494a0. [DOI] [PubMed] [Google Scholar]
  9. Dunn T. M., Hahn S., Ogden S., Schleif R. F. An operator at -280 base pairs that is required for repression of araBAD operon promoter: addition of DNA helical turns between the operator and promoter cyclically hinders repression. Proc Natl Acad Sci U S A. 1984 Aug;81(16):5017–5020. doi: 10.1073/pnas.81.16.5017. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Ebbole D. J., Zalkin H. Detection of pur operon-attenuated mRNA and accumulated degradation intermediates in Bacillus subtilis. J Biol Chem. 1988 Aug 5;263(22):10894–10902. [PubMed] [Google Scholar]
  11. 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]
  12. 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]
  13. Fritz H. J., Bicknäse H., Gleumes B., Heibach C., Rosahl S., Ehring R. Characterization of two mutations in the Escherichia coli galE gene inactivating the second galactose operator and comparative studies of repressor binding. EMBO J. 1983;2(12):2129–2135. doi: 10.1002/j.1460-2075.1983.tb01713.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Gilbert W., Maxam A. The nucleotide sequence of the lac operator. Proc Natl Acad Sci U S A. 1973 Dec;70(12):3581–3584. doi: 10.1073/pnas.70.12.3581. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Gillespie D., Spiegelman S. A quantitative assay for DNA-RNA hybrids with DNA immobilized on a membrane. J Mol Biol. 1965 Jul;12(3):829–842. doi: 10.1016/s0022-2836(65)80331-x. [DOI] [PubMed] [Google Scholar]
  16. 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]
  17. Gralla J. D. Bacterial gene regulation from distant DNA sites. Cell. 1989 Apr 21;57(2):193–195. doi: 10.1016/0092-8674(89)90955-0. [DOI] [PubMed] [Google Scholar]
  18. Gunsalus R. P., Yanofsky C. Nucleotide sequence and expression of Escherichia coli trpR, the structural gene for the trp aporepressor. Proc Natl Acad Sci U S A. 1980 Dec;77(12):7117–7121. doi: 10.1073/pnas.77.12.7117. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Haber R., Adhya S. Interaction of spatially separated protein-DNA complexes for control of gene expression: operator conversions. Proc Natl Acad Sci U S A. 1988 Dec;85(24):9683–9687. doi: 10.1073/pnas.85.24.9683. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Hasunuma K., Sekiguchi M. Effect of dna mutations on the replication of plasmid pSC101 in Escherichia coli K-12. J Bacteriol. 1979 Mar;137(3):1095–1099. doi: 10.1128/jb.137.3.1095-1099.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. 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]
  22. He B., Shiau A., Choi K. Y., Zalkin H., Smith J. M. Genes of the Escherichia coli pur regulon are negatively controlled by a repressor-operator interaction. J Bacteriol. 1990 Aug;172(8):4555–4562. doi: 10.1128/jb.172.8.4555-4562.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Hochschild A., Ptashne M. Cooperative binding of lambda repressors to sites separated by integral turns of the DNA helix. Cell. 1986 Mar 14;44(5):681–687. doi: 10.1016/0092-8674(86)90833-0. [DOI] [PubMed] [Google Scholar]
  24. 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]
  25. Hsieh W. T., Whitson P. A., Matthews K. S., Wells R. D. Influence of sequence and distance between two operators on interaction with the lac repressor. J Biol Chem. 1987 Oct 25;262(30):14583–14591. [PubMed] [Google Scholar]
  26. Huo L., Martin K. J., Schleif R. Alternative DNA loops regulate the arabinose operon in Escherichia coli. Proc Natl Acad Sci U S A. 1988 Aug;85(15):5444–5448. doi: 10.1073/pnas.85.15.5444. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Irani M. H., Orosz L., Adhya S. A control element within a structural gene: the gal operon of Escherichia coli. Cell. 1983 Mar;32(3):783–788. doi: 10.1016/0092-8674(83)90064-8. [DOI] [PubMed] [Google Scholar]
  28. Kelley R. L., Yanofsky C. Trp aporepressor production is controlled by autogenous regulation and inefficient translation. Proc Natl Acad Sci U S A. 1982 May;79(10):3120–3124. doi: 10.1073/pnas.79.10.3120. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. 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]
  30. Krämer H., Niemöller M., Amouyal M., Revet B., von Wilcken-Bergmann B., Müller-Hill B. lac repressor forms loops with linear DNA carrying two suitably spaced lac operators. EMBO J. 1987 May;6(5):1481–1491. doi: 10.1002/j.1460-2075.1987.tb02390.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Kunkel T. A., Roberts J. D., Zakour R. A. Rapid and efficient site-specific mutagenesis without phenotypic selection. Methods Enzymol. 1987;154:367–382. doi: 10.1016/0076-6879(87)54085-x. [DOI] [PubMed] [Google Scholar]
  32. 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]
  33. Masters M. Location of purine genes on the physical map of Escherichia coli. J Bacteriol. 1990 Mar;172(3):1173–1173. doi: 10.1128/jb.172.3.1173.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. 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]
  35. 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]
  36. Ptashne M. Gene regulation by proteins acting nearby and at a distance. Nature. 1986 Aug 21;322(6081):697–701. doi: 10.1038/322697a0. [DOI] [PubMed] [Google Scholar]
  37. Reznikoff W. S., Siegele D. A., Cowing D. W., Gross C. A. The regulation of transcription initiation in bacteria. Annu Rev Genet. 1985;19:355–387. doi: 10.1146/annurev.ge.19.120185.002035. [DOI] [PubMed] [Google Scholar]
  38. Reznikoff W. S., Winter R. B., Hurley C. K. The location of the repressor binding sites in the lac operon. Proc Natl Acad Sci U S A. 1974 Jun;71(6):2314–2318. doi: 10.1073/pnas.71.6.2314. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. 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]
  40. Rolfes R. J., Zalkin H. Purification of the Escherichia coli purine regulon repressor and identification of corepressors. J Bacteriol. 1990 Oct;172(10):5637–5642. doi: 10.1128/jb.172.10.5637-5642.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. 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]
  42. Sampei G., Mizobuchi K. The organization of the purL gene encoding 5'-phosphoribosylformylglycinamide amidotransferase of Escherichia coli. J Biol Chem. 1989 Dec 15;264(35):21230–21238. [PubMed] [Google Scholar]
  43. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. 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]
  45. Shapira S. K., Chou J., Richaud F. V., Casadaban M. J. New versatile plasmid vectors for expression of hybrid proteins coded by a cloned gene fused to lacZ gene sequences encoding an enzymatically active carboxy-terminal portion of beta-galactosidase. Gene. 1983 Nov;25(1):71–82. doi: 10.1016/0378-1119(83)90169-5. [DOI] [PubMed] [Google Scholar]
  46. 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]
  47. Tiedeman A. A., DeMarini D. J., Parker J., Smith J. M. DNA sequence of the purC gene encoding 5'-phosphoribosyl-5-aminoimidazole-4-N-succinocarboxamide synthetase and organization of the dapA-purC region of Escherichia coli K-12. J Bacteriol. 1990 Oct;172(10):6035–6041. doi: 10.1128/jb.172.10.6035-6041.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. 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]
  49. 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]
  50. 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]
  51. 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]
  52. 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]
  53. 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]
  54. 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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