Skip to main content
SpringerLink
Log in

DNA repair properties of Escherichia coli tif-1, recAo281 and lexA1 strains deficient in single-strand DNA binding protein

  • Published:
Molecular and General Genetics MGG Aims and scope Submit manuscript

Summary

Mutations affecting single-strand DNA binding protein (SSB) impair induction of mutagenic (SOS) repair. To further investigate the role of SSB in SOS induction and DNA repair, isogenic strains were constructed combining the ssb +, ssb-1 or ssb-113 alleles with one or more mutations known to alter regulation of damage inducible functions. As is true in ssb + strains tif-1 (recA441) was found to allow thermal induction of prophage λ+ and Weigle reactivation in ssb-1 and ssb-113 strains. Furthermore, tif-1 decreased the UV sensitivity of the ssb-113 strain slightly and permitted UV induction of prophage λ+ at 30°C. Strains carrying the recAo281 allele were also constructed. This mutation causes high constitutive levels of RecA protein synthesis and relieves much of the UV sensitivity conferred by lexA − alleles without restoring SOS (error-prone) repair. In contrast, the recAo281 allele failed to alleviate the UV sensitivity associated with either ssb − mutation. In a lexA1 recAo281 background the ssb-1 mutation increased the extent of postirradiation DNA degradation and concommitantly increased UV sensitivity 20-fold to the level exhibited by a recA1 strain. The ssb-113 mutation also increased UV sensitivity markedly in this background but did so without greatly increasing postirradiation DNA degradation. These results suggest a direct role for SSB in recombinational repair apart from and in addition to its role in facilitating induction of the recA-lexA regulon.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Bagg A, Kenyon CJ, Walker GC (1981) Inducibility of a gene product required for UV and chemical mutagenesis in Escherichia coli. Proc Natl Acad Sci USA 78:5789–5753

    Google Scholar 

  • Baluch J, Chase JW, Sussman R (1980) Synthesis of recA protein and induction of bacteriophage lambda in single-strand deoxyribonucleic acid-binding protein mutants of Escherichia coli. J Bacteriol 144:489–498

    Google Scholar 

  • Brent R, Ptashne M (1980) The lexA gene product represses its own promoter. Proc Natl Acad Sci USA 77:1932–1936

    Google Scholar 

  • Casaregola S, D'Ari R, Huisman O (1982) Quantitative evaluation of recA gene expression in Escherichia coli. Mol Gen Genet 185:430–439

    Google Scholar 

  • Castellazzi M, George J, Buttin G (1972) Prophage induction and cell division in E. coli. I. Further characterization of the thermo-sensitive mutation tif-1 whose expression mimics the effect of UV irradiation. Mol Gen Genet 119:139–152

    Google Scholar 

  • Chase JW, Masker WE (1977) Deoxyribonucleic acid repair in Escherichia coli mutants deficient in the 5′ → 3′ exonuclease activity of deoxyribonucleic acid polymerase I and exonuclease-VII. J Bacteriol 130:667–675

    Google Scholar 

  • Chase JW, Murphy JB, Whittier RF, Lorensen E, Sninsky JJ (1983) Amplification of ssb-1 mutant single-stranded DNA-binding protein in Escherichia coli. J Mol Biol 163:in press

  • Chrysogelos S, Griffith J (1982) E. coli single-strand binding protein organizes single-stranded DNA in nucleosome-like units. Proc Natl Acad Sci USA 79:5803–5807

    Google Scholar 

  • Cox MM, Lehman IR (1981) recA protein of Escherichia coli promotes branch migration, a kinetically distinct phase of DNA strand exchange. Proc Natl Acad Sci USA 78:3433–3437

    Google Scholar 

  • Craig NL, Roberts JW (1980) E. coli recA protein-directed cleavage of phage λ repressor requires polynucleotide. Nature 283:26–30

    Google Scholar 

  • Flory J, Radding CM (1982) Visualization of RecA protein with DNA: a priming effect of single-strand-binding protein. Cell 28:747–756

    Google Scholar 

  • Ganesan AK, Seawell PC (1975) The effect of lexA and recF mutations on post-replicational repair and DNA synthesis in Escherichia coli K-12. Mol Gen Genet 141:189–205

    Google Scholar 

  • Golub EI, Low KB (1983) Indirect stimulation of genetic recombination. Proc Natl Acad Sci USA 80, in press

  • Huisman O, D'Ari R (1981) An inducible DNA replication-cell division coupling mechanism in E. coli. Nature 290:797–799

    Google Scholar 

  • Johnson BF (1977) Genetic mapping of the lexC-113 mutation. Mol Gen Genet 157:91–97

    Google Scholar 

  • Kenyon CJ, Walker GC (1980) DNA-damaging agents stimulate gene expression at specific loci in Escherichia coli. Proc Natl Acad Sci USA 77:2819–2823

    Google Scholar 

  • Kenyon CJ, Walker GC (1981) Expression of the E. coli uvrA gene is inducible. Nature 289:808–810

    Google Scholar 

  • Kirby EP, Jacob F, Goldthwait DA (1967) Prophage induction and filament formation in a mutant strain of Escherichia coli. Proc Natl Acad Sci USA 58:1903–1910

    Google Scholar 

  • Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 277:680–685

    Google Scholar 

  • Lieberman HB, Witkin EM (1981) Variable expression of the ssb-1 allele in different strains of Escherichia coli K12 and B: differential suppression of its effects on DNA replication, DNA repair and ultraviolet mutagenesis. Mol Gen Genet 183:348–355

    Google Scholar 

  • Lieberman HB, Witkin EM (1983) 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 190:92–100

    Google Scholar 

  • Little JW, Harper JE (1979) Identification of the lexA gene product of Escherichia coli K-12. Proc Natl Acad Sci USA 76:6147–6151

    Google Scholar 

  • Little JW, Edmiston SH, Pacelli LZ, Mount DW (1980) Cleavage of the Escherichia coli lexA protein by the recA protease. Proc Natl Acad Sci USA 77:3225–3229

    Google Scholar 

  • Little JW, Mount DW, Yanisch-Perron CR (1981) Purified lexA protein is a repressor of the recA and lexA genes. Proc Natl Acad Sci USA 78:4199–4203

    Google Scholar 

  • Little JW, Mount DW (1982) The SOS regulatory system of Escherichia coli. Cell 29:11–22

    Google Scholar 

  • Livneh Z, Lehman IR (1982) Recombinational bypass of pyrimidine dimers promoted by the recA protein of Escherichia coli. Proc Natl Acad Sci USA 79:3171–3175

    Google Scholar 

  • Mackay V, Linn S (1976) Selective inhibition of the DNase activity of the recBC enzyme by the DNA binding protein from Escherichia coli. J Biol Chem 251:3716–3719

    Google Scholar 

  • McEntee K, Weinstock GM, Lehman IR (1980) recA protein-catalyzed strand assimilation: stimulation by Escherichia coli single-stranded DNA-binding protein. Proc Natl Acad Sci USA 77:857–861

    Google Scholar 

  • McEntee K, Weinstock GM (1981) tif-1 mutation alters polynucleotide recognition by the recA protein of Escherichia coli. Proc Natl Acad Sci USA 78:6061–6065

    Google Scholar 

  • Miller J (1972) Experiments in molecular genetics. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York

    Google Scholar 

  • Molineux IJ, Gefter ML (1975) Properties of the Escherichia coli DNA-binding (unwinding) protein interaction with nucleolytic enzymes and DNA. J Mol Biol 98:811–825

    Google Scholar 

  • Phizicky EM, Roberts JW (1981) Induction of SOS functions: regulation of proteolytic activity of E. coli RecA protein by interaction with DNA and nucleoside triphosphate. Cell 25:259–267

    Google Scholar 

  • Resnick J, Sussman R (1982) Escherichia coli single-strand DNA binding protein from wild type and lexC113 mutant affects in vitro proteolytic cleavage of phage λ repressor. Proc Natl Acad Sci USA 79:2832–2835

    Google Scholar 

  • Roberts JW, Roberts CW, Craig NL (1978) Escherichia coli recA gene product inactivates phage λ repressor. Proc Natl Acad Sci USA 75:4714–4718

    Google Scholar 

  • Sancar A, Williams KR, Chase JW, Rupp WD (1981) Sequences of the ssb gene and protein. Proc Natl Acad Sci USA 78:4274–4278

    Google Scholar 

  • Sancar GB, Sancar A, Little JW, Rupp WD (1982) The uvrB gene of Escherichia coli has both lexA-repressed and lexA-independent promoters. Cell 28:523–530

    Google Scholar 

  • Uhlin BE, Volkert RM, Clark AJ, Sancar A, Rupp WD (1982) Nucleotide sequence of a recA operator mutation. Mol Gen Genet 185:251–254

    Google Scholar 

  • Vales LD, Chase JW, Murphy JB (1980) Effect of ssbA1 and lexC113 mutations on lambda prophage induction, bacteriophage growth, and cell survival. J Bacteriol 143:887–896

    Google Scholar 

  • Volkert MR, George DL, Witkin EM (1976) Partial suppression of the LexA phenotype by mutations (rnm) which restore ultraviolet resistance but not ultraviolet mutability to Escherichia coli B/r uvr A LexA. Mutat Res 36:17–28

    Google Scholar 

  • Volkert MR, Spencer DF, Clark AJ (1979) Indirect and intragenic suppression of the lexA102 mutation in E. coli B/r. Mol Gen Genet 177:129–137

    Google Scholar 

  • Volkert MR, Margossian LJ, Clark AJ (1981) Evidence that rnmB is the operator of the Escherichia coli recA gene. Proc Natl Acad Sci USA 78:1786–1790

    Google Scholar 

  • Wang T-c V, Smith KC (1982) Effects of the ssb-1 and ssb-113 mutations on survival and DNA repair in UV-irradiated uvrB strains of Escherichia coli K-12. J Bacteriol 151:186–192

    Google Scholar 

  • Weigle JJ (1953) Induction of mutations in a bacterial virus. Proc Natl Acad Sci USA 39:628–636

    Google Scholar 

  • Weinstock GM, McEntee K, Lehman IR (1979) ATP-dependent renaturation of DNA catalyzed by the recA protein of Escherichia coli. Proc Natl Acad Sci USA 76:126–130

    Google Scholar 

  • Weinstock GM, McEntee K (1981) RecA protein dependent proteolysis of bacteriophage λ repressor. Characterization of the reaction and stimulation by DNA-binding proteins. J Biol Chem 256:10883–10888

    Google Scholar 

  • West SC, Cassuto E, Howard-Flanders P (1981) Mechanism of E. coli RecA protein directed strand exchanges in post-replication repair of DNA. Nature 294:659–662

    Google Scholar 

  • Whittier RF, Chase JW (1981) DNA repair in E. coli strains deficient in single-strand DNA binding protein. Mol Gen Genet 183:341–347

    Google Scholar 

  • Williams KR, L'Italien JJ, Sillerud L, Spicer E, Chase JW, Konigsberg W (1982a) Mutant DNA binding proteins: HPLC peptide mapping and physicochemical properties. Fed Proc 41:1198

    Google Scholar 

  • Williams KR, L'Italien JJ, Guggenheimer RA, Sillerud L, Spicer E, Chase JW, Konigsberg W (1982b) Comparative peptide mapping by HPLC: identification of single amino acid substitutions in temperature sensitive mutants. Proc of the IV International Conference on Methods in Protein Sequence Analysis, Humana Press, Clifton, New Jersey, pp 499–507

    Google Scholar 

  • Williams KR, Spicer EK, LoPresti MB, Guggenheimer RA, Chase JW (1983) Limited proteolysis studies on the Escherichia coli single-strand DNA binding protein: evidence for a functionally homologous domain in both the Escherichia coli and T4 DNA binding proteins. J Biol Chem 258, in press

  • Witkin EM (1976) Ultraviolet mutagenesis and inducible DNA repair in Escherichia coli. Bacteriol Rev 40:869–907

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Molecular Biology, Albert Einstein College of Medicine, 1300 Morris Park Avenue, 10461, Bronx, New York, USA

    Robert F. Whittier & John W. Chase

Authors
  1. Robert F. Whittier
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. John W. Chase
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

Communicated by B.A. Bridges

Rights and permissions

Reprints and permissions

About this article

Cite this article

Whittier, R.F., Chase, J.W. DNA repair properties of Escherichia coli tif-1, recAo281 and lexA1 strains deficient in single-strand DNA binding protein. Mol Gen Genet 190, 101–111 (1983). https://doi.org/10.1007/BF00330330

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00330330

Keywords

Search

Navigation