Skip to main content
SpringerLink
Log in

PsiB polypeptide prevents activation of RecA protein in Escherichia coli

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

Summary

We further characterize a novel plasmid function preventing SOS induction called Psi (Plasmid SOS Inhibition). We show that Psi function is expressed by psiB, a gene located at coordinate 54.9 of plasmid R6-5 and near oriT, the origin of conjugal transfer. Deletions and amber mutations of the psiB gene permitted us to demonstrate that PsiB polypeptide (apparent molecular weight, 12 kDa) is responsible for Psi function. PsiB protein prevents recA730-promoted mutagenesis and intra-chromosomal recombination but not recombination following conjugation. Overproduction of PsiB protein sensitizes the host cell to UV irradiation. We propose that PsiB polypeptide has an anti-SOS action by inhibiting activation of RecA protein, thus preventing the occurrence of LexA-controlled functions.

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

  • Andoh T, Ozeki H (1968) Suppressor gene su3 + of E. coli, a structural gene for tyrosine tRNA. Proc Natl Acad Sci USA 59:792–798

    Google Scholar 

  • Bachmann BJ (1972) Pedigrees of some mutant strains of Escherichia coli K12. Bacteriol Rev 36:525–557

    Google Scholar 

  • Bagdasarian M, Bailone A, Bagdasarian MM, Manning PA, Lurz R, Timmis KN, Devoret R (1986) An inhibitor of SOS induction specified by a plasmid locus in Escherichia coli Proc Natl Acad Sci USA 83:5723–5726

    Google Scholar 

  • Burckhardt SE, Woodgate R, Scheuermann RH, Echols H (1988) The UmuD mutagenesis protein of E. coli: overproduction, purification and cleavage by RecA. Proc Natl. Acad Sci USA 85:1811–1815

    Google Scholar 

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

    Google Scholar 

  • Chang ACY, Cohen SN (1978) Construction and characterization of amplifiable multicopy DNA cloning vehicules derived from p15A cryptic miniplasmid. J Bacteriol 134:1141–1156

    Google Scholar 

  • Chase JW, Merril BM, Williams KR (1983) F sex factor encodes a single-stranded DNA binding protein (SSB) with extensive sequence homology to Escherichia coli SSB. Proc Natl Acad Sci USA 80:5480–5484

    Google Scholar 

  • Clark AJ, Sandler SJ, Willis KD, Chu C, Blanar MA, Lovett ST (1984) Genes of the RecE and RecF pathways of conjugational recombination in E. coli. Cold Spring Harbor Symp Quant Biol 49:453–462

    Google Scholar 

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

    Google Scholar 

  • Craig NL, Roberts JW (1981) Function of nucleoside triphosphate and polynucleotide in Escherichia coli recA protein-directed cleavage of phage lambda repressor. J Biol Chem 256:8039–8044

    Google Scholar 

  • Devoret R, Blanco M (1970) Mutants of Escherichia coli K12 (λ)+ non-inducible by thymine deprivation. I. Method of isolation and classes of mutants obtained. Mol Gen Genet 107:272–280

    Google Scholar 

  • Devoret R, Pierre M, Moreau PL (1983) Prophage Ø80 is induced in Escherichia coli K12 recA430. Mol Gen Genet 189:199–206

    Google Scholar 

  • Devoret R, Bailone A, Dutreix M, Moreau PL, Sommer S, Bagdasarian M (1988) Regulation of activation of RecA protein in E. coli. In: Friedberg EC, Hanawalt PC (eds) UCLA Symposia on Molecular and Cellular Biology: Mechanisms and Consequences of DNA Damage Processing. Liss Inc, New York

    Google Scholar 

  • Fürste JP, Pansegrau W, Frank R, Blocker H, Scholz P, Bagdasarian M, Lanka E (1986) Molecular cloning of the RP4 DNA primase region in a multi-host-range tacP expression vector. Gene 48:119–131

    Google Scholar 

  • Golub EI, Low KB (1986a) Derepression of single-stranded DNA-binding protein genes on plasmids derepressed for conjugation and complementation of an E. coli ssb - mutation by these genes. Mol Gen Genet 204:410–416

    Google Scholar 

  • Golub EI, Low KB (1986b) Unrelated conjugative plasmids have sequences which are homologous to the leading region of the F factor. J Bacteriol 166:670–672

    Google Scholar 

  • Hershfield V, Boyer HW, Chow L, Helinski DR (1976) Characterization of a mini-ColE1 plasmid. J Bacteriol 126:447–453

    Google Scholar 

  • Howard-Flanders P, West SC, Stasiak A (1984) Role of RecA protein spiral filaments in genetic recombination. Nature 309:215–220

    Google Scholar 

  • Huisman O, D'Ari R (1983) Effect of suppressors of SOS-mediated filamentation of sfiA operon in Escherichia coli. J Bacteriol 153:169–175

    Google Scholar 

  • Humphreys GO, Willshaw GA, Smith HR, Anderson ES (1976) Mutagenesis of plasmid DNA with hydroxylamine: Isolation of mutants of multi-copy plasmids. Mol Gen Genet 145:101–108

    Google Scholar 

  • Irino N, Nakayama K, Nakayama H (1986) The recQ gene of Escherichia coli K12: Primary structure and evidence for SOS regulation. Mol Gen Genet 205:298–304

    Google Scholar 

  • Kolodkin AL, Capage MA, Golub EI, Low KB (1983) F sex factor of Escherichia coli K12 codes for a single-stranded DNA binding protein. Proc Natl Acad Sci USA 80:4422–4426

    Google Scholar 

  • Konrad EB, Lehman IR (1975) Novel mutants of Escherichia coli that accumulate very small DNA replicative intermediates. Proc Natl Acad Sci USA 72:2150–2154

    Google Scholar 

  • Little JW (1984) Auto-digestion of lexA and phage lambda repressors. Proc Natl Acad USA 81:1375–1379

    Google Scholar 

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

    Google Scholar 

  • Lu C, Echols H (1987) RecA protein and SOS. Correlation of mutagenesis phenotype with binding of mutant RecA proteins to duplex DNA and LexA cleavage. J Mol Biol 196:497–504

    Google Scholar 

  • Marsh L, Walker GC (1987) New phenotypes associated with mucAB: Alteration of a mucA sequence homologous to the LexA cleavage site. J Bacteriol 169:1818–1823

    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 

  • Meyer RR, Voegele DW, Ruben SM, Rein DC, Trela JM (1982) Influence of single-stranded DNA-binding protein on recA induction in Escherichia coli. Mutat Res 94:299–313

    Google Scholar 

  • Moreau PL (1987) Effects of overproduction of single-stranded DNA binding protein on RecA-protein-dependent processes in Escherichia coli. J Mol Biol 194:621–634

    Google Scholar 

  • Mount DW (1977) A mutant of Escherichia coli showing constitutive expression of the lysogenic induction and error-prone DNA repair pathways. Proc Natl Acad Sci USA 74:300–304

    Google Scholar 

  • Nohmi T, Battista JR, Dodson LA, Walker GC (1988) RecA-mediated cleavage activates UmuD for mutagenesis: Mechanistic relationship between transcriptional derepression and post-translational activation. Proc Natl Acad Sci USA 85:1816–1820

    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 

  • Picksley SM, Lloyd RG, Buckman C (1984) Genetic analysis and regulation of inducible recombination in Escherichia coli K12. Cold Spring Harbor Symp Quant Biol 49:469–474

    Google Scholar 

  • Quillardet P, Moreau PL, Ginsburg H, Mount DW, Devoret R (1982) Cell survival, UV reactivation and induction of prophage lambda in Escherichia coli K12 overproducing RecA protein. Mol Gen Genet 188:37–43

    Google Scholar 

  • Radding CM, Flory J, Wu A, Kahn R, DasGupta C, Gonda D, Bianchi M, Tsang SS (1983a) Three phases in homologous pairing: Polymerisation of recA on single-stranded DNA, synapsis, and polar strand exchange. Cold Spring Harbor Symp Quant Biol 47:821–828

    Google Scholar 

  • Radding CM, DasGupta C, Wu AM, Kahn R, Flory J (1983b) Recombination activities of Escherichia coli RecA protein: Synapsis and strand exchange. Nucleic Acids Res: 535–548

  • radman M (1975) SOS repair hypothesis: Phenomenology of an inducible DNA repair which is accompanied by mutagenesis. In: Hanawalt PC, Setlow RB (eds) Molecular Mechanisms for Repair of DNA. Plenum Press, New York, pp 355–367

    Google Scholar 

  • Sancar A, Hack AM, Rupp WD (1979) Simple method for identification of plasmid-coded proteins. J Bacteriol 137:692–693

    Google Scholar 

  • Sharp PA, Cohen SN, Davidson N (1973) Electron microscope heteroduplex studies of sequence relations among plamids of Escherichia coli. II. Structure of drug resistance (R) factors and F factors. J Mol Biol 75:235–255

    Google Scholar 

  • Shinagawa H, Iwasaki H, Kato T, Nakata A (1988) RecA protein-dependent cleavage of UmuD protein and SOS mutagenesis. Proc Natl Acad Sci USA 85:1806–1810

    Google Scholar 

  • Slilaty SN, Little JW (1987) Lysine-156 and serine-119 are required for LexA repressor cleavage: A possible mechanism. Proc Natl Acad Sci USA 84:3987–3991

    Google Scholar 

  • Smith GR (1988) Homologous recombination in procaryotes. Microbiol Rev 52:1–28

    Google Scholar 

  • Sommer S, Bailone A, Devoret R (1985) SOS induction by thermosensitive replication mutants of miniF plasmid. Mol Gen Genet 198:456–464

    Google Scholar 

  • Strike P, Lodwick D (1988) Sequence and further characterization of the imp operon (I-group mutation and protection) of the plasmid TP110. J Cell Biochem [Suppl] 12 A:326

    Google Scholar 

  • Timmis KN, Cabello F, Cohen SN (1978) Cloning and characterization of EcoRI and HindIII restriction endonuclease-generated fragments of antibiotic resistance plasmid R6-5 and R6. Mol Gen Genet 162:121–137

    Google Scholar 

  • Wang WB, Tessman ES (1985) Evidence that the recA441 (tif) mutant of Escherichia coli K12 contains a thermosensitive intragenic suppressor of RecA constitutive protease activity. J Bacteriol 163:407–409

    Google Scholar 

  • Wanner BL, Wieder S, McSharry R (1981) Use of bacteriophage transposon Mu d1 to determine the orientation for three proC-linked phosphate starvation inducible (psi) genes in Escherichia coli K12. J Bacteriol 146:93–101

    Google Scholar 

  • Willetts N, Wilkins B (1984) Processing of plasmid DNA during bacterial conjugation. Microbiol Rev 48:24–41

    Google Scholar 

  • Witkin EM (1982) From Gainesville to Toulouse: The evolution of a model. Biochimie (Paris) 64:549–555

    Google Scholar 

  • Witkin EM, Kogoma T (1984) Involvement of the activated form of RecA protein in SOS mutagenesis and stable DNA replication in Escherichia coli. Proc Natl Acad Sci USA 81:7539–7543

    Google Scholar 

  • Womble DD, Taylor DP, Rownd RH (1977) Method for obtaining more accurate covalently closed circular plasmid to chromosome ratios from bacterial lysates by dye buoyant density centrifugation. J Bacteriol 130:148–153

    Google Scholar 

  • Zieg J, Maples VF, Kushner SR (1978) Recombination levels of Escherichia coli K12 mutants deficient in various replication, recombination, or repair genes. J Bacteriol 134:958–966

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. G.E.M.C., Enzymologie, Centre National de la Recherche Scientifique, F-91190, Gif-sur-Y vette, France

    Adriana Bailone, Suzanne Sommer, Jérome Célérier & Raymond Devoret

  2. Michigan Biotechnology Institute, 48909, Lansing, MI, USA

    Assar Bäckman, Mira M. Bagdasarian & Michael Bagdasarian

Authors
  1. Adriana Bailone
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Assar Bäckman
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Suzanne Sommer
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Jérome Célérier
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Mira M. Bagdasarian
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Michael Bagdasarian
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Raymond Devoret
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

Communicated by G.R. Smith

Rights and permissions

Reprints and permissions

About this article

Cite this article

Bailone, A., Bäckman, A., Sommer, S. et al. PsiB polypeptide prevents activation of RecA protein in Escherichia coli . Mol Gen Genet 214, 389–395 (1988). https://doi.org/10.1007/BF00330471

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00330471

Key words

Search

Navigation