Skip to main content
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1985 May;82(10):3325–3329. doi: 10.1073/pnas.82.10.3325

Dual role for Escherichia coli RecA protein in SOS mutagenesis.

D G Ennis, B Fisher, S Edmiston, D W Mount
PMCID: PMC397768  PMID: 3159017

Abstract

Induction of the Escherichia coli SOS system increases the ability of the cells to perform DNA repair and mutagenesis. Previous work has shown that this increased mutagenesis is the result of derepression of specific genes through a complex regulatory mechanism controlled by LexA and RecA proteins. One role of RecA protein in this process is to facilitate proteolytic cleavage of LexA protein (the repressor) in response to an inducing signal that reversibly activates RecA protein to perform this function. We show that activated RecA protein plays a second role in SOS mutagenesis, as revealed by analyzing repair of UV-damaged phage lambda in host mutants with alterations in the SOS regulatory system. First, phage mutagenesis was not expressed constitutively in a mutant that is derepressed through lack of functional LexA protein; activated RecA protein was still required. Second, phage mutagenesis was constitutively expressed in the presence of recA mutations that alter RecA protein so that it is activated in normally growing cells. There was also RecA-dependent constitutive expression of SOS mutagenesis in host mutants that lack functional LexA protein and carry plasmids. We discuss several possible biochemical mechanisms for this second role of activated RecA protein in SOS mutagenesis.

Full text

PDF

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bagg A., Kenyon C. J., Walker G. C. Inducibility of a gene product required for UV and chemical mutagenesis in Escherichia coli. Proc Natl Acad Sci U S A. 1981 Sep;78(9):5749–5753. doi: 10.1073/pnas.78.9.5749. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Blanco M., Herrera G., Collado P., Rebollo J. E., Botella L. M. Influence of RecA protein on induced mutagenesis. Biochimie. 1982 Aug-Sep;64(8-9):633–636. doi: 10.1016/s0300-9084(82)80102-8. [DOI] [PubMed] [Google Scholar]
  3. Capage M. A., Scott J. R. SOS induction by P1 Km miniplasmids. J Bacteriol. 1983 Aug;155(2):473–480. doi: 10.1128/jb.155.2.473-480.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Devoret R., Pierre M., Moreau P. L. Prophage phi 80 is induced in Escherichia coli K12 recA430. Mol Gen Genet. 1983;189(2):199–206. doi: 10.1007/BF00337804. [DOI] [PubMed] [Google Scholar]
  5. Dressler D., Potter H. Molecular mechanisms in genetic recombination. Annu Rev Biochem. 1982;51:727–761. doi: 10.1146/annurev.bi.51.070182.003455. [DOI] [PubMed] [Google Scholar]
  6. Echols H., Lu C., Burgers P. M. Mutator strains of Escherichia coli, mutD and dnaQ, with defective exonucleolytic editing by DNA polymerase III holoenzyme. Proc Natl Acad Sci U S A. 1983 Apr;80(8):2189–2192. doi: 10.1073/pnas.80.8.2189. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Echols H. Mutation rate: some biological and biochemical considerations. Biochimie. 1982 Aug-Sep;64(8-9):571–575. doi: 10.1016/s0300-9084(82)80089-8. [DOI] [PubMed] [Google Scholar]
  8. Elledge S. J., Walker G. C. Proteins required for ultraviolet light and chemical mutagenesis. Identification of the products of the umuC locus of Escherichia coli. J Mol Biol. 1983 Feb 25;164(2):175–192. doi: 10.1016/0022-2836(83)90074-8. [DOI] [PubMed] [Google Scholar]
  9. Fersht A. R., Knill-Jones J. W. Contribution of 3' leads to 5' exonuclease activity of DNA polymerase III holoenzyme from Escherichia coli to specificity. J Mol Biol. 1983 Apr 25;165(4):669–682. doi: 10.1016/s0022-2836(83)80273-3. [DOI] [PubMed] [Google Scholar]
  10. Ginsburg H., Edmiston S. H., Harper J., Mount D. W. Isolation and characterization of an operator-constitutive mutation in the recA gene of E. coli K-12. Mol Gen Genet. 1982;187(1):4–11. doi: 10.1007/BF00384376. [DOI] [PubMed] [Google Scholar]
  11. Kahn M., Kolter R., Thomas C., Figurski D., Meyer R., Remaut E., Helinski D. R. Plasmid cloning vehicles derived from plasmids ColE1, F, R6K, and RK2. Methods Enzymol. 1979;68:268–280. doi: 10.1016/0076-6879(79)68019-9. [DOI] [PubMed] [Google Scholar]
  12. Krueger J. H., Elledge S. J., Walker G. C. Isolation and characterization of Tn5 insertion mutations in the lexA gene of Escherichia coli. J Bacteriol. 1983 Mar;153(3):1368–1378. doi: 10.1128/jb.153.3.1368-1378.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Lackey D., Krauss S. W., Linn S. Isolation of an altered form of DNA polymerase I from Escherichia coli cells induced for recA/lexA functions. Proc Natl Acad Sci U S A. 1982 Jan;79(2):330–334. doi: 10.1073/pnas.79.2.330. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Lieberman H. B., Witkin E. M. DNA degradation, UV sensitivity and SOS-mediated mutagenesis in strains of Escherichia coli deficient in single-strand DNA binding protein: effects of mutations and treatments that alter levels of Exonuclease V or recA protein. Mol Gen Genet. 1983;190(1):92–100. doi: 10.1007/BF00330329. [DOI] [PubMed] [Google Scholar]
  15. Little J. W. Autodigestion of lexA and phage lambda repressors. Proc Natl Acad Sci U S A. 1984 Mar;81(5):1375–1379. doi: 10.1073/pnas.81.5.1375. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Little J. W., Mount D. W. The SOS regulatory system of Escherichia coli. Cell. 1982 May;29(1):11–22. doi: 10.1016/0092-8674(82)90085-x. [DOI] [PubMed] [Google Scholar]
  17. McCann J., Spingarn N. E., Kobori J., Ames B. N. Detection of carcinogens as mutagens: bacterial tester strains with R factor plasmids. Proc Natl Acad Sci U S A. 1975 Mar;72(3):979–983. doi: 10.1073/pnas.72.3.979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Miller J. H. Mutational specificity in bacteria. Annu Rev Genet. 1983;17:215–238. doi: 10.1146/annurev.ge.17.120183.001243. [DOI] [PubMed] [Google Scholar]
  19. Miura A., Tomizawa J. Mutation and recombination of bacteriophage lambda: effect of ultraviolet radiation. Proc Natl Acad Sci U S A. 1970 Dec;67(4):1722–1726. doi: 10.1073/pnas.67.4.1722. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Morand P., Blanco M., Devoret R. Characterization of lexB mutations in Escherichia coli K-12. J Bacteriol. 1977 Aug;131(2):572–582. doi: 10.1128/jb.131.2.572-582.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Mount D. W. A mutant of Escherichia coli showing constitutive expression of the lysogenic induction and error-prone DNA repair pathways. Proc Natl Acad Sci U S A. 1977 Jan;74(1):300–304. doi: 10.1073/pnas.74.1.300. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Mount D. W., Harris A. W., Fuerst C. R., Siminovitch L. Mutations in bacteriophage lambda affecting particle morphogenesis. Virology. 1968 May;35(1):134–149. doi: 10.1016/0042-6822(68)90313-9. [DOI] [PubMed] [Google Scholar]
  23. Radding C. M. Homologous pairing and strand exchange in genetic recombination. Annu Rev Genet. 1982;16:405–437. doi: 10.1146/annurev.ge.16.120182.002201. [DOI] [PubMed] [Google Scholar]
  24. Roberts J. W., Roberts C. W. Two mutations that alter the regulatory activity of E. coli recA protein. Nature. 1981 Apr 2;290(5805):422–424. doi: 10.1038/290422a0. [DOI] [PubMed] [Google Scholar]
  25. Scheuermann R., Tam S., Burgers P. M., Lu C., Echols H. Identification of the epsilon-subunit of Escherichia coli DNA polymerase III holoenzyme as the dnaQ gene product: a fidelity subunit for DNA replication. Proc Natl Acad Sci U S A. 1983 Dec;80(23):7085–7089. doi: 10.1073/pnas.80.23.7085. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Schultz D. W., Taylor A. F., Smith G. R. Escherichia coli RecBC pseudorevertants lacking chi recombinational hotspot activity. J Bacteriol. 1983 Aug;155(2):664–680. doi: 10.1128/jb.155.2.664-680.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Shinagawa H., Kato T., Ise T., Makino K., Nakata A. Cloning and characterization of the umu operon responsible for inducible mutagenesis in Escherichia coli. Gene. 1983 Aug;23(2):167–174. doi: 10.1016/0378-1119(83)90048-3. [DOI] [PubMed] [Google Scholar]
  28. Smith G. R. Deletion mutations of the immunity region of coliphage lambda. Virology. 1975 Apr;64(2):544–552. doi: 10.1016/0042-6822(75)90131-2. [DOI] [PubMed] [Google Scholar]
  29. 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]
  30. Weigle J. J. Induction of Mutations in a Bacterial Virus. Proc Natl Acad Sci U S A. 1953 Jul;39(7):628–636. doi: 10.1073/pnas.39.7.628. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Winans S. C., Walker G. C. Genetic localization and characterization of a pKM101-coded endonuclease. J Bacteriol. 1983 Jun;154(3):1117–1125. doi: 10.1128/jb.154.3.1117-1125.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Witkin E. M., Kogoma T. Involvement of the activated form of RecA protein in SOS mutagenesis and stable DNA replication in Escherichia coli. Proc Natl Acad Sci U S A. 1984 Dec;81(23):7539–7543. doi: 10.1073/pnas.81.23.7539. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Witkin E. M., McCall J. O., Volkert M. R., Wermundsen I. E. Constitutive expression of SOS functions and modulation of mutagenesis resulting from resolution of genetic instability at or near the recA locus of Escherichia coli. Mol Gen Genet. 1982;185(1):43–50. doi: 10.1007/BF00333788. [DOI] [PubMed] [Google Scholar]
  34. Witkin E. M. Ultraviolet mutagenesis and inducible DNA repair in Escherichia coli. Bacteriol Rev. 1976 Dec;40(4):869–907. doi: 10.1128/br.40.4.869-907.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Witkin E. M., Wermundsen I. E. Targeted and untargeted mutagenesis by various inducers of SOS functions in Escherichia coli. Cold Spring Harb Symp Quant Biol. 1979;43(Pt 2):881–886. doi: 10.1101/sqb.1979.043.01.095. [DOI] [PubMed] [Google Scholar]
  36. Wood R. D., Hutchinson F. Non-targeted mutagenesis of unirradiated lambda phage in Escherichia coli host cells irradiated with ultraviolet light. J Mol Biol. 1984 Mar 5;173(3):293–305. doi: 10.1016/0022-2836(84)90122-0. [DOI] [PubMed] [Google Scholar]
  37. Wood R. D., Skopek T. R., Hutchinson F. Changes in DNA base sequence induced by targeted mutagenesis of lambda phage by ultraviolet light. J Mol Biol. 1984 Mar 5;173(3):273–291. doi: 10.1016/0022-2836(84)90121-9. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES