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. 1987 Aug;169(8):3435–3440. doi: 10.1128/jb.169.8.3435-3440.1987

Regulation of the Escherichia coli uvrD gene in vivo.

H M Arthur, D R Cavanagh, P W Finch, P T Emmerson
PMCID: PMC212414  PMID: 3038838

Abstract

The roles of two putative promoter sequences, P1 and P2, and a potential antiterminator sequence found in the uvrD control region were examined in vivo. Constitutive and SOS-induced levels of uvrD mRNA were determined by S1 mapping, and it was shown that the majority of uvrD transcripts are from P1, while P2 plays only a minor role. A series of increasing deletions from the 5' end of the uvrD gene was used to assay transcription in the promoterless vector pKO-1. Loss of just the -35 region of P1 was sufficient to switch off detectable transcription from both P1 and P2. Disruption of the antiterminator by site-specific mutagenesis had no effect on constitutive levels of transcription, but led to a significant increase over wild-type levels following SOS induction. This suggests that the attenuator comes into play following DNA damage to moderate the increase in UvrD protein synthesis.

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Selected References

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  1. Abdel-Monem M., Chanal M. C., Hoffmann-Berling H. DNA unwinding enzyme II of Escherichia coli. 1. Purification and characterization of the ATPase activity. Eur J Biochem. 1977 Sep 15;79(1):33–38. doi: 10.1111/j.1432-1033.1977.tb11780.x. [DOI] [PubMed] [Google Scholar]
  2. Abdel-Monem M., Dürwald H., Hoffmann-Berling H. DNA unwinding enzyme II of Escherichia coli. 2. Characterization of the DNA unwinding activity. Eur J Biochem. 1977 Sep 15;79(1):39–45. doi: 10.1111/j.1432-1033.1977.tb11781.x. [DOI] [PubMed] [Google Scholar]
  3. Arthur H. M., Bramhill D., Eastlake P. B., Emmerson P. T. Cloning of the uvrD gene of E. coli and identification of the product. Gene. 1982 Oct;19(3):285–295. doi: 10.1016/0378-1119(82)90018-x. [DOI] [PubMed] [Google Scholar]
  4. Arthur H. M., Eastlake P. B. Transcriptional control of the uvrD gene of Escherichia coli. Gene. 1983 Nov;25(2-3):309–316. doi: 10.1016/0378-1119(83)90235-4. [DOI] [PubMed] [Google Scholar]
  5. Arthur H. M., Lloyd R. G. Hyper-recombination in uvrD mutants of Escherichia coli K-12. Mol Gen Genet. 1980;180(1):185–191. doi: 10.1007/BF00267368. [DOI] [PubMed] [Google Scholar]
  6. 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]
  7. 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]
  8. Caron P. R., Kushner S. R., Grossman L. Involvement of helicase II (uvrD gene product) and DNA polymerase I in excision mediated by the uvrABC protein complex. Proc Natl Acad Sci U S A. 1985 Aug;82(15):4925–4929. doi: 10.1073/pnas.82.15.4925. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Easton A. M., Kushner S. R. Transcription of the uvrD gene of Escherichia coli is controlled by the lexA repressor and by attenuation. Nucleic Acids Res. 1983 Dec 20;11(24):8625–8640. doi: 10.1093/nar/11.24.8625. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Finch P. W., Emmerson P. T. The nucleotide sequence of the uvrD gene of E. coli. Nucleic Acids Res. 1984 Jul 25;12(14):5789–5799. doi: 10.1093/nar/12.14.5789. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Finch P., Emmerson P. T. Nucleotide sequence of the regulatory region of the uvrD gene of Escherichia coli. Gene. 1983 Nov;25(2-3):317–323. doi: 10.1016/0378-1119(83)90236-6. [DOI] [PubMed] [Google Scholar]
  12. HOWARD-FLANDERS P., THERIOT L. A method for selecting radiation-sensitive mutants of Escherichia coli. Genetics. 1962 Sep;47:1219–1224. doi: 10.1093/genetics/47.9.1219. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. 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]
  14. Hickson I. D., Arthur H. M., Bramhill D., Emmerson P. T. The E. coli uvrD gene product is DNA helicase II. Mol Gen Genet. 1983;190(2):265–270. doi: 10.1007/BF00330649. [DOI] [PubMed] [Google Scholar]
  15. Howard-Flanders P., Bardwell E. Effects of recB21, recF143, and uvrD152 on recombination in lambda bacteriophage-prophage and Hfr by F- crosses. J Bacteriol. 1981 Nov;148(2):739–743. doi: 10.1128/jb.148.2.739-743.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Husain I., Van Houten B., Thomas D. C., Abdel-Monem M., Sancar A. Effect of DNA polymerase I and DNA helicase II on the turnover rate of UvrABC excision nuclease. Proc Natl Acad Sci U S A. 1985 Oct;82(20):6774–6778. doi: 10.1073/pnas.82.20.6774. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Kuemmerle N. B., Masker W. E. Effect of the uvrD mutation on excision repair. J Bacteriol. 1980 May;142(2):535–546. doi: 10.1128/jb.142.2.535-546.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Kumura K., Sekiguchi M. Identification of the uvrD gene product of Escherichia coli as DNA helicase II and its induction by DNA-damaging agents. J Biol Chem. 1984 Feb 10;259(3):1560–1565. [PubMed] [Google Scholar]
  19. Kumura K., Sekiguchi M., Steinum A. L., Seeberg E. Stimulation of the UvrABC enzyme-catalyzed repair reactions by the UvrD protein (DNA helicase II). Nucleic Acids Res. 1985 Mar 11;13(5):1483–1492. doi: 10.1093/nar/13.5.1483. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Lloyd R. G. lexA dependent recombination in uvrD strains of Escherichia coli. Mol Gen Genet. 1983;189(1):157–161. doi: 10.1007/BF00326069. [DOI] [PubMed] [Google Scholar]
  21. Lu A. L., Clark S., Modrich P. Methyl-directed repair of DNA base-pair mismatches in vitro. Proc Natl Acad Sci U S A. 1983 Aug;80(15):4639–4643. doi: 10.1073/pnas.80.15.4639. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Maples V. F., Kushner S. R. DNA repair in Escherichia coli: identification of the uvrD gene product. Proc Natl Acad Sci U S A. 1982 Sep;79(18):5616–5620. doi: 10.1073/pnas.79.18.5616. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. McDonell M. W., Simon M. N., Studier F. W. Analysis of restriction fragments of T7 DNA and determination of molecular weights by electrophoresis in neutral and alkaline gels. J Mol Biol. 1977 Feb 15;110(1):119–146. doi: 10.1016/s0022-2836(77)80102-2. [DOI] [PubMed] [Google Scholar]
  24. 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]
  25. Messing J., Crea R., Seeburg P. H. A system for shotgun DNA sequencing. Nucleic Acids Res. 1981 Jan 24;9(2):309–321. doi: 10.1093/nar/9.2.309. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Oeda K., Horiuchi T., Sekiguchi M. Molecular cloning of the uvrD gene of Escherichia coli that controls ultraviolet sensitivity and spontaneous mutation frequency. Mol Gen Genet. 1981;184(2):191–199. doi: 10.1007/BF00272904. [DOI] [PubMed] [Google Scholar]
  27. Oeda K., Horiuchi T., Sekiguchi M. The uvrD gene of E. coli encodes a DNA-dependent ATPase. Nature. 1982 Jul 1;298(5869):98–100. doi: 10.1038/298098a0. [DOI] [PubMed] [Google Scholar]
  28. Rothman R. H., Clark A. J. Defective excision and postreplication repair of UV-damaged DNA in a recL mutant strain of E. coli K-12. Mol Gen Genet. 1977 Oct 24;155(3):267–277. doi: 10.1007/BF00272805. [DOI] [PubMed] [Google Scholar]
  29. 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]
  30. Siegel E. C. The Escherichia coli uvrD gene is inducible by DNA damage. Mol Gen Genet. 1983;191(3):397–400. doi: 10.1007/BF00425753. [DOI] [PubMed] [Google Scholar]
  31. Siegel E. C. Ultraviolet-sensitive mutator strain of Escherichia coli K-12. J Bacteriol. 1973 Jan;113(1):145–160. doi: 10.1128/jb.113.1.145-160.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Smirnov G. B., Filkova E. V., Skavronskaya A. G. The mutator property of uvr502 mutation affecting UV sensitivity of Escherichia coli. Mol Gen Genet. 1972;118(1):51–56. doi: 10.1007/BF02428332. [DOI] [PubMed] [Google Scholar]
  33. Taucher-Scholz G., Hoffmann-Berling H. Identification of the gene for DNA helicase II of Escherichia coli. Eur J Biochem. 1983 Dec 15;137(3):573–580. doi: 10.1111/j.1432-1033.1983.tb07864.x. [DOI] [PubMed] [Google Scholar]
  34. Tinoco I., Jr, Borer P. N., Dengler B., Levin M. D., Uhlenbeck O. C., Crothers D. M., Bralla J. Improved estimation of secondary structure in ribonucleic acids. Nat New Biol. 1973 Nov 14;246(150):40–41. doi: 10.1038/newbio246040a0. [DOI] [PubMed] [Google Scholar]
  35. Van Sluis C. A., Mattern I. E., Paterson M. C. Properties of uvrE mutants of Escherichia coli K12. I. Effects of UV irradiation on DNA metabolism. Mutat Res. 1974 Dec;25(3):273–279. doi: 10.1016/0027-5107(74)90055-4. [DOI] [PubMed] [Google Scholar]
  36. 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]
  37. Yamamoto Y., Ogawa T., Shinagawa H., Nakayama T., Matsuo H., Ogawa H. Determination of the initiation sites of transcription and translation of the uvrD gene of Escherichia coli. J Biochem. 1986 Jun;99(6):1579–1590. doi: 10.1093/oxfordjournals.jbchem.a135631. [DOI] [PubMed] [Google Scholar]
  38. Zoller M. J., Smith M. Oligonucleotide-directed mutagenesis of DNA fragments cloned into M13 vectors. Methods Enzymol. 1983;100:468–500. doi: 10.1016/0076-6879(83)00074-9. [DOI] [PubMed] [Google Scholar]
  39. van den Elzen P. J., Konings R. N., Veltkamp E., Nijkamp H. J. Transcription of bacteriocinogenic plasmid CloDF13 in vivo and in vitro: structure of the cloacin immunity operon. J Bacteriol. 1980 Nov;144(2):579–591. doi: 10.1128/jb.144.2.579-591.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]

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