Skip to main content
NCBI home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1993 Feb;175(3):642–646. doi: 10.1128/jb.175.3.642-646.1993

Characterization of dinY, a new Escherichia coli DNA repair gene whose products are damage inducible even in a lexA(Def) background.

C Petit 1, C Cayrol 1, C Lesca 1, P Kaiser 1, C Thompson 1, M Defais 1
PMCID: PMC196200  PMID: 8423139

Abstract

Bacteriophage Mu dX(Ap lac) was used to isolate a mutation in an Escherichia coli lexA(Def) strain representing a previously undescribed gene (dinY) which does not seem to be under the direct control of LexA. The insertion created a dinY::lacZ fusion in which beta-galactosidase expression required a DNA-damaging treatment (UV irradiation or mitomycin) and activable RecA protein. This strain showed a decreased Weigle reactivation of bacteriophage lambda. However, it was fully inducible for UV mutagenesis. Two-dimensional gel electrophoresis analysis identified two spots absent in the mutant which were both UV inducible only in the presence of activated RecA protein (RecA*). This finding suggests that the dinY::lacZ fusion lies in a gene either that is under the direct control of activated RecA or whose product undergoes RecA*-dependent posttranscriptional/posttranslational modification(s). The dinY gene may also control the expression of some other gene(s) and/or lie in an operon. The fusion was mapped at a position between 41 and 41.5 min on the E. coli chromosome, in the vicinity of the ruv operon.

Full text

PDF
642

Images in this article

644Image
on p.644

Selected References

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

  1. Baker T. A., Howe M. M., Gross C. A. Mu dX, a derivative of Mu d1 (lac Apr) which makes stable lacZ fusions at high temperature. J Bacteriol. 1983 Nov;156(2):970–974. doi: 10.1128/jb.156.2.970-974.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bonner C. A., Randall S. K., Rayssiguier C., Radman M., Eritja R., Kaplan B. E., McEntee K., Goodman M. F. Purification and characterization of an inducible Escherichia coli DNA polymerase capable of insertion and bypass at abasic lesions in DNA. J Biol Chem. 1988 Dec 15;263(35):18946–18952. [PubMed] [Google Scholar]
  3. Burckhardt S. E., Woodgate R., Scheuermann R. H., Echols H. UmuD mutagenesis protein of Escherichia coli: overproduction, purification, and cleavage by RecA. Proc Natl Acad Sci U S A. 1988 Mar;85(6):1811–1815. doi: 10.1073/pnas.85.6.1811. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Calsou P., Villaverde A., Defais M. Activated RecA protein may induce expression of a gene that is not controlled by the LexA repressor and whose function is required for mutagenesis and repair of UV-irradiated bacteriophage lambda. J Bacteriol. 1987 Oct;169(10):4816–4821. doi: 10.1128/jb.169.10.4816-4821.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Crowl R. M., Boyce R. P., Echols H. Repressor cleavage as a prophage induction mechanism: hypersensitivity of a mutant lambda cI protein to recA-mediated proteolysis. J Mol Biol. 1981 Nov 15;152(4):815–819. doi: 10.1016/0022-2836(81)90128-5. [DOI] [PubMed] [Google Scholar]
  6. Donnelly C. E., Walker G. C. groE mutants of Escherichia coli are defective in umuDC-dependent UV mutagenesis. J Bacteriol. 1989 Nov;171(11):6117–6125. doi: 10.1128/jb.171.11.6117-6125.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Dutreix M., Moreau P. L., Bailone A., Galibert F., Battista J. R., Walker G. C., Devoret R. New recA mutations that dissociate the various RecA protein activities in Escherichia coli provide evidence for an additional role for RecA protein in UV mutagenesis. J Bacteriol. 1989 May;171(5):2415–2423. doi: 10.1128/jb.171.5.2415-2423.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Ennis D. G., Fisher B., Edmiston S., Mount D. W. Dual role for Escherichia coli RecA protein in SOS mutagenesis. Proc Natl Acad Sci U S A. 1985 May;82(10):3325–3329. doi: 10.1073/pnas.82.10.3325. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Garrels J. I. The QUEST system for quantitative analysis of two-dimensional gels. J Biol Chem. 1989 Mar 25;264(9):5269–5282. [PubMed] [Google Scholar]
  10. Goloubinoff P., Gatenby A. A., Lorimer G. H. GroE heat-shock proteins promote assembly of foreign prokaryotic ribulose bisphosphate carboxylase oligomers in Escherichia coli. Nature. 1989 Jan 5;337(6202):44–47. doi: 10.1038/337044a0. [DOI] [PubMed] [Google Scholar]
  11. Hochstrasser D. F., Patchornik A., Merril C. R. Development of polyacrylamide gels that improve the separation of proteins and their detection by silver staining. Anal Biochem. 1988 Sep;173(2):412–423. doi: 10.1016/0003-2697(88)90208-4. [DOI] [PubMed] [Google Scholar]
  12. Kato T., Nakano E. Effects of the umuC36 mutation on ultraviolet-radiation-induced base-change and frameshift mutations in Escherichia coli. Mutat Res. 1981 Oct;83(3):307–319. doi: 10.1016/0027-5107(81)90014-2. [DOI] [PubMed] [Google Scholar]
  13. Kenyon C. J., Walker G. C. DNA-damaging agents stimulate gene expression at specific loci in Escherichia coli. Proc Natl Acad Sci U S A. 1980 May;77(5):2819–2823. doi: 10.1073/pnas.77.5.2819. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kogoma T., Farr S. B., Joyce K. M., Natvig D. O. Isolation of gene fusions (soi::lacZ) inducible by oxidative stress in Escherichia coli. Proc Natl Acad Sci U S A. 1988 Jul;85(13):4799–4803. doi: 10.1073/pnas.85.13.4799. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. 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]
  16. Krueger J. H., Walker G. C. groEL and dnaK genes of Escherichia coli are induced by UV irradiation and nalidixic acid in an htpR+-dependent fashion. Proc Natl Acad Sci U S A. 1984 Mar;81(5):1499–1503. doi: 10.1073/pnas.81.5.1499. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Lesca C., Petit C., Defais M. UV induction of LexA independent proteins which could be involved in SOS repair. Biochimie. 1991 Apr;73(4):407–409. doi: 10.1016/0300-9084(91)90107-c. [DOI] [PubMed] [Google Scholar]
  18. 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]
  19. Little J. W. The SOS regulatory system: control of its state by the level of RecA protease. J Mol Biol. 1983 Jul 15;167(4):791–808. doi: 10.1016/s0022-2836(83)80111-9. [DOI] [PubMed] [Google Scholar]
  20. Liu S. K., Tessman I. groE genes affect SOS repair in Escherichia coli. J Bacteriol. 1990 Oct;172(10):6135–6138. doi: 10.1128/jb.172.10.6135-6138.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Lu C., Scheuermann R. H., Echols H. Capacity of RecA protein to bind preferentially to UV lesions and inhibit the editing subunit (epsilon) of DNA polymerase III: a possible mechanism for SOS-induced targeted mutagenesis. Proc Natl Acad Sci U S A. 1986 Feb;83(3):619–623. doi: 10.1073/pnas.83.3.619. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Maenhaut-Michel G., Caillet-Fauquet P. Genetic control of the UV-induced SOS mutator effect in single- and double-stranded DNA phages. Mutat Res. 1990 Jun;230(2):241–254. doi: 10.1016/0027-5107(90)90062-9. [DOI] [PubMed] [Google Scholar]
  23. 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]
  24. Mount D. W., Little J. W., Edmiston S. H. Influence of plasmids carrying the lexA gene on DNA repair and related processes in Escherichia coli K-12. Mol Gen Genet. 1980 Feb;177(3):477–483. doi: 10.1007/BF00271487. [DOI] [PubMed] [Google Scholar]
  25. Nohmi T., Battista J. R., Dodson L. A., Walker G. C. RecA-mediated cleavage activates UmuD for mutagenesis: mechanistic relationship between transcriptional derepression and posttranslational activation. Proc Natl Acad Sci U S A. 1988 Mar;85(6):1816–1820. doi: 10.1073/pnas.85.6.1816. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. O'Farrell P. H. High resolution two-dimensional electrophoresis of proteins. J Biol Chem. 1975 May 25;250(10):4007–4021. [PMC free article] [PubMed] [Google Scholar]
  27. Sedgwick S. G., Yarranton G. T., Heath R. W. Lysogenic induction of lambdoid phages in lexA mutants of Escherichia coli. Mol Gen Genet. 1981;184(3):457–459. doi: 10.1007/BF00352522. [DOI] [PubMed] [Google Scholar]
  28. Shinagawa H., Iwasaki H., Kato T., Nakata A. RecA protein-dependent cleavage of UmuD protein and SOS mutagenesis. Proc Natl Acad Sci U S A. 1988 Mar;85(6):1806–1810. doi: 10.1073/pnas.85.6.1806. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Sweasy J. B., Witkin E. M., Sinha N., Roegner-Maniscalco V. RecA protein of Escherichia coli has a third essential role in SOS mutator activity. J Bacteriol. 1990 Jun;172(6):3030–3036. doi: 10.1128/jb.172.6.3030-3036.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. 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]
  31. 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]
  32. 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]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES