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. 1987 May 26;15(10):4273–4289. doi: 10.1093/nar/15.10.4273

Regulation of the Escherichia coli excision repair gene uvrC. Overlap between the uvrC structural gene and the region coding for a 24 kD protein.

G F Moolenaar, C A van Sluis, C Backendorf, P van de Putte
PMCID: PMC340847  PMID: 3295776

Abstract

The UvrA, UvrB and UvrC proteins of E. coli are subunits of a DNA repair enzyme, the ABC exonuclease. In this paper we study the uvrC regulatory region. The uvrC structural gene is preceded by an open reading frame encoding a 24 kD protein. A uvrC promoter has been mapped within this gene. The transcription start of a second promoter located 5' of the 24 kD gene is mapped in vivo. We show that transcription from both promoters on the chromosome is not inducible by UV damage. The possible translation start codons of the UvrC and of the 24 kD protein are determined. Sequences encoding the N-terminal part of the UvrC protein overlap with sequences encoding the C-terminal part of the 24 kD protein. To examine a possible function of the 24 kD gene in repair, a 24 kD insertion mutant was created in the chromosome. The mutant however only slightly affects the UV sensitivity of the cell. Transcription of P3 alone provides sufficient UvrC protein for the normal repair of UV lesions.

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Selected References

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  1. Alton N. K., Vapnek D. Nucleotide sequence analysis of the chloramphenicol resistance transposon Tn9. Nature. 1979 Dec 20;282(5741):864–869. doi: 10.1038/282864a0. [DOI] [PubMed] [Google Scholar]
  2. Backendorf C., Brandsma J. A., Kartasova T., van de Putte P. In vivo regulation of the uvrA gene: role of the "-10" and "-35" promoter regions. Nucleic Acids Res. 1983 Sep 10;11(17):5795–5810. doi: 10.1093/nar/11.17.5795. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Berk A. J., Sharp P. A. Sizing and mapping of early adenovirus mRNAs by gel electrophoresis of S1 endonuclease-digested hybrids. Cell. 1977 Nov;12(3):721–732. doi: 10.1016/0092-8674(77)90272-0. [DOI] [PubMed] [Google Scholar]
  4. Brandsma J. A., Bosch D., de Ruÿter M., van de Putte P. Analysis of the regulatory region of the ssb gene of Escherichia coli. Nucleic Acids Res. 1985 Jul 25;13(14):5095–5109. doi: 10.1093/nar/13.14.5095. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Das A., Yanofsky C. A ribosome binding site sequence is necessary for efficient expression of the distal gene of a translationally-coupled gene pair. Nucleic Acids Res. 1984 Jun 11;12(11):4757–4768. doi: 10.1093/nar/12.11.4757. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Fogliano M., Schendel P. F. Evidence for the inducibility of the uvrB operon. Nature. 1981 Jan 15;289(5794):196–198. doi: 10.1038/289196a0. [DOI] [PubMed] [Google Scholar]
  7. Forster J. W., Strike P. Organisation and control of the Escherichia coli uvrC gene. Gene. 1985;35(1-2):71–82. doi: 10.1016/0378-1119(85)90159-3. [DOI] [PubMed] [Google Scholar]
  8. Granger-Schnarr M., Schnarr M., van Sluis C. A. In vitro study of the interaction of the LexA repressor and the UvrC protein with a uvrC regulatory region. FEBS Lett. 1986 Mar 17;198(1):61–65. doi: 10.1016/0014-5793(86)81185-1. [DOI] [PubMed] [Google Scholar]
  9. Gren E. J. Recognition of messenger RNA during translational initiation in Escherichia coli. Biochimie. 1984 Jan;66(1):1–29. doi: 10.1016/0300-9084(84)90188-3. [DOI] [PubMed] [Google Scholar]
  10. Howard-Flanders P., Boyce R. P., Theriot L. Three loci in Escherichia coli K-12 that control the excision of pyrimidine dimers and certain other mutagen products from DNA. Genetics. 1966 Jun;53(6):1119–1136. doi: 10.1093/genetics/53.6.1119. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Kenyon C. J., Walker G. C. Expression of the E. coli uvrA gene is inducible. Nature. 1981 Feb 26;289(5800):808–810. doi: 10.1038/289808a0. [DOI] [PubMed] [Google Scholar]
  12. Maxam A. M., Gilbert W. Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol. 1980;65(1):499–560. doi: 10.1016/s0076-6879(80)65059-9. [DOI] [PubMed] [Google Scholar]
  13. McKenney K., Shimatake H., Court D., Schmeissner U., Brady C., Rosenberg M. A system to study promoter and terminator signals recognized by Escherichia coli RNA polymerase. Gene Amplif Anal. 1981;2:383–415. [PubMed] [Google Scholar]
  14. Ohtsubo H., Ohtsubo E. Nucleotide sequence of an insertion element, IS1. Proc Natl Acad Sci U S A. 1978 Feb;75(2):615–619. doi: 10.1073/pnas.75.2.615. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Reddy P., Peterkofsky A., McKenney K. Translational efficiency of the Escherichia coli adenylate cyclase gene: mutating the UUG initiation codon to GUG or AUG results in increased gene expression. Proc Natl Acad Sci U S A. 1985 Sep;82(17):5656–5660. doi: 10.1073/pnas.82.17.5656. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Sharma S., Stark T. F., Beattie W. G., Moses R. E. Multiple control elements for the uvrC gene unit of Escherichia coli. Nucleic Acids Res. 1986 Mar 11;14(5):2301–2318. doi: 10.1093/nar/14.5.2301. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  18. Stüber D., Bujard H. Organization of transcriptional signals in plasmids pBR322 and pACYC184. Proc Natl Acad Sci U S A. 1981 Jan;78(1):167–171. doi: 10.1073/pnas.78.1.167. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Walker G. C. Inducible DNA repair systems. Annu Rev Biochem. 1985;54:425–457. doi: 10.1146/annurev.bi.54.070185.002233. [DOI] [PubMed] [Google Scholar]
  20. Yanisch-Perron C., Vieira J., Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. doi: 10.1016/0378-1119(85)90120-9. [DOI] [PubMed] [Google Scholar]
  21. Yoakum G. H., Grossman L. Identification of E. coli uvrC protein. Nature. 1981 Jul 9;292(5819):171–173. doi: 10.1038/292171a0. [DOI] [PubMed] [Google Scholar]
  22. de Boer H. A., Comstock L. J., Vasser M. The tac promoter: a functional hybrid derived from the trp and lac promoters. Proc Natl Acad Sci U S A. 1983 Jan;80(1):21–25. doi: 10.1073/pnas.80.1.21. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. de Wind N., de Jong M., Meijer M., Stuitje A. R. Site-directed mutagenesis of the Escherichia coli chromosome near oriC: identification and characterization of asnC, a regulatory element in E. coli asparagine metabolism. Nucleic Acids Res. 1985 Dec 20;13(24):8797–8811. doi: 10.1093/nar/13.24.8797. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. van Sluis C. A., Moolenaar G. F., Backendorf C. Regulation of the uvrC gene of Escherichia coli K12: localization and characterization of a damage-inducible promoter. EMBO J. 1983;2(12):2313–2318. doi: 10.1002/j.1460-2075.1983.tb01739.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. van den Berg E. A., Geerse R. H., Pannekoek H., van de Putte P. In vivo transcription of the E. coli uvrB gene: both promoters are inducible by UV. Nucleic Acids Res. 1983 Jul 11;11(13):4355–4363. doi: 10.1093/nar/11.13.4355. [DOI] [PMC free article] [PubMed] [Google Scholar]

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