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. 1985 Oct;164(1):276–281. doi: 10.1128/jb.164.1.276-281.1985

Analysis of the ruv locus of Escherichia coli K-12 and identification of the gene product.

P V Attfield, F E Benson, R G Lloyd
PMCID: PMC214240  PMID: 2995311

Abstract

The ruv gene of Escherichia coli, which is associated with inducible mechanisms of DNA repair and recombination, has been cloned into the low-copy plasmid vector pHSG415. The recombinant plasmid pPVA101 fully complements the DNA repair-deficient phenotype of ruv mutants. Restriction endonuclease analysis of this plasmid revealed a 10.6-kilobase (kb) HindIII DNA insert which contained a 7.7-kb PstI fragment identified as being from the chromosomal ruv region. Deletion analysis and Tn1000 insertional inactivation of ruv function located the ruv coding region to a 2.2-kb section of the cloned DNA fragment. A comparison of the proteins encoded by ruv wild-type and mutant plasmids identified the gene product as a protein of molecular weight 41,000.

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Selected References

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  1. 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]
  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. Cohen S. N., Chang A. C., Hsu L. Nonchromosomal antibiotic resistance in bacteria: genetic transformation of Escherichia coli by R-factor DNA. Proc Natl Acad Sci U S A. 1972 Aug;69(8):2110–2114. doi: 10.1073/pnas.69.8.2110. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. George J., Castellazzi M., Buttin G. Prophage induction and cell division in E. coli. III. Mutations sfiA and sfiB restore division in tif and lon strains and permit the expression of mutator properties of tif. Mol Gen Genet. 1975 Oct 22;140(4):309–332. [PubMed] [Google Scholar]
  5. Grindley N. D. Transposition of Tn3 and related transposons. Cell. 1983 Jan;32(1):3–5. doi: 10.1016/0092-8674(83)90490-7. [DOI] [PubMed] [Google Scholar]
  6. Guyer M. S. The gamma delta sequence of F is an insertion sequence. J Mol Biol. 1978 Dec 15;126(3):347–365. doi: 10.1016/0022-2836(78)90045-1. [DOI] [PubMed] [Google Scholar]
  7. Hashimoto-Gotoh T., Franklin F. C., Nordheim A., Timmis K. N. Specific-purpose plasmid cloning vectors. I. Low copy number, temperature-sensitive, mobilization-defective pSC101-derived containment vectors. Gene. 1981 Dec;16(1-3):227–235. doi: 10.1016/0378-1119(81)90079-2. [DOI] [PubMed] [Google Scholar]
  8. Horii Z., Clark A. J. Genetic analysis of the recF pathway to genetic recombination in Escherichia coli K12: isolation and characterization of mutants. J Mol Biol. 1973 Oct 25;80(2):327–344. doi: 10.1016/0022-2836(73)90176-9. [DOI] [PubMed] [Google Scholar]
  9. 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]
  10. Ish-Horowicz D., Burke J. F. Rapid and efficient cosmid cloning. Nucleic Acids Res. 1981 Jul 10;9(13):2989–2998. doi: 10.1093/nar/9.13.2989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Iyehara H., Otsuji N. Ultraviolet light sensitivity of Escherichia coli K-12 strains carrying ruv mutations in combination with uvrA or lon mutant alleles. J Bacteriol. 1975 Feb;121(2):735–736. doi: 10.1128/jb.121.2.735-736.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Kleckner N. Transposable elements in prokaryotes. Annu Rev Genet. 1981;15:341–404. doi: 10.1146/annurev.ge.15.120181.002013. [DOI] [PubMed] [Google Scholar]
  13. Little J. W., Mount D. W., Yanisch-Perron C. R. Purified lexA protein is a repressor of the recA and lexA genes. Proc Natl Acad Sci U S A. 1981 Jul;78(7):4199–4203. doi: 10.1073/pnas.78.7.4199. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Lloyd R. G., Barbour S. D. The genetic location of the sbcA gene of Escherichia coli. Mol Gen Genet. 1974;134(2):157–171. doi: 10.1007/BF00268417. [DOI] [PubMed] [Google Scholar]
  15. Lloyd R. G., Benson F. E., Shurvinton C. E. Effect of ruv mutations on recombination and DNA repair in Escherichia coli K12. Mol Gen Genet. 1984;194(1-2):303–309. doi: 10.1007/BF00383532. [DOI] [PubMed] [Google Scholar]
  16. Lloyd R. G., Low B., Godson G. N., Birge E. A. Isolation and characterization of an Escherichia coli K-12 mutant with a temperature-sensitive recA- phenotype. J Bacteriol. 1974 Oct;120(1):407–415. doi: 10.1128/jb.120.1.407-415.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Lloyd R. G., Low B. Some genetic consequences of changes in the level of recA gene function in Escherichia coli K-12. Genetics. 1976 Dec;84(4):675–695. doi: 10.1093/genetics/84.4.675. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Lloyd R. G., Picksley S. M., Prescott C. Inducible expression of a gene specific to the RecF pathway for recombination in Escherichia coli K12. Mol Gen Genet. 1983;190(1):162–167. doi: 10.1007/BF00330340. [DOI] [PubMed] [Google Scholar]
  19. Lovett S. T., Clark A. J. Genetic analysis of regulation of the RecF pathway of recombination in Escherichia coli K-12. J Bacteriol. 1983 Mar;153(3):1471–1478. doi: 10.1128/jb.153.3.1471-1478.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Lovett S. T., Clark A. J. Genetic analysis of the recJ gene of Escherichia coli K-12. J Bacteriol. 1984 Jan;157(1):190–196. doi: 10.1128/jb.157.1.190-196.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Nakayama H., Nakayama K., Nakayama R., Irino N., Nakayama Y., Hanawalt P. C. Isolation and genetic characterization of a thymineless death-resistant mutant of Escherichia coli K12: identification of a new mutation (recQ1) that blocks the RecF recombination pathway. Mol Gen Genet. 1984;195(3):474–480. doi: 10.1007/BF00341449. [DOI] [PubMed] [Google Scholar]
  22. Norrander J., Kempe T., Messing J. Construction of improved M13 vectors using oligodeoxynucleotide-directed mutagenesis. Gene. 1983 Dec;26(1):101–106. doi: 10.1016/0378-1119(83)90040-9. [DOI] [PubMed] [Google Scholar]
  23. Otsuji N., Iyehara-Ogawa H. Thermoresistant revertants of an Escherichia coli strain carrying tif-1 and ruv mutations: non-suppressibility of ruv by sfi. J Bacteriol. 1979 Apr;138(1):1–6. doi: 10.1128/jb.138.1.1-6.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Otsuji N., Iyehara H., Hideshima Y. Isolation and characterization of an Escherichia coli ruv mutant which forms nonseptate filaments after low doses of ultraviolet light irradiation. J Bacteriol. 1974 Feb;117(2):337–344. doi: 10.1128/jb.117.2.337-344.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Picksley S. M., Attfield P. V., Lloyd R. G. Repair of DNA double-strand breaks in Escherichia coli K12 requires a functional recN product. Mol Gen Genet. 1984;195(1-2):267–274. doi: 10.1007/BF00332758. [DOI] [PubMed] [Google Scholar]
  26. Reed R. R. Transposon-mediated site-specific recombination: a defined in vitro system. Cell. 1981 Sep;25(3):713–719. doi: 10.1016/0092-8674(81)90178-1. [DOI] [PubMed] [Google Scholar]
  27. Sancar A., Wharton R. P., Seltzer S., Kacinski B. M., Clarke N. D., Rupp W. D. Identification of the uvrA gene product. J Mol Biol. 1981 May 5;148(1):45–62. doi: 10.1016/0022-2836(81)90234-5. [DOI] [PubMed] [Google Scholar]
  28. Shurvinton C. E., Lloyd R. G., Benson F. E., Attfield P. V. Genetic analysis and molecular cloning of the Escherichia coli ruv gene. Mol Gen Genet. 1984;194(1-2):322–329. doi: 10.1007/BF00383535. [DOI] [PubMed] [Google Scholar]
  29. Shurvinton C. E., Lloyd R. G. Damage to DNA induces expression of the ruv gene of Escherichia coli. Mol Gen Genet. 1982;185(2):352–355. doi: 10.1007/BF00330811. [DOI] [PubMed] [Google Scholar]
  30. Vieira J., Messing J. The pUC plasmids, an M13mp7-derived system for insertion mutagenesis and sequencing with synthetic universal primers. Gene. 1982 Oct;19(3):259–268. doi: 10.1016/0378-1119(82)90015-4. [DOI] [PubMed] [Google Scholar]
  31. Walker G. C. Mutagenesis and inducible responses to deoxyribonucleic acid damage in Escherichia coli. Microbiol Rev. 1984 Mar;48(1):60–93. doi: 10.1128/mr.48.1.60-93.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. West S. C., Emmerson P. T. Induction of protein synthesis in Escherichia coli following UV- or gamma-irradiation, mitomycin C treatment or tif Expression. Mol Gen Genet. 1977 Feb 28;151(1):57–67. doi: 10.1007/BF00446913. [DOI] [PubMed] [Google Scholar]

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