Summary
DNA is more extensively degraded after it is damaged in recA mutants of E. coli than in wild type cells. All data presented here are consistent with the recA gene product, protein X, being an inhibitor of nalidixic acid induced degradation of the bulk DNA (but not of newly replicated DNA). Production of protein X also is correlated with appearance of various “S.O.S.” repair functions. Evidence was obtained by comparing the rates of protein X synthesis and solubilization of uniformly-labeled DNA in intact cells, incubated in the presence of nalidixic acid. A set of mutants at the lexA locus produced protein X at different rates and degraded their DNA at rates which were inversely correlated to their rates of protein X production. A low concentration of rifampicin quite specifically inhibited protein X production by wild type E. coli, and allowed more rapid DNA degradation. After the DNA was damaged by the incubation of cells in the presence of nalidixic acid, cells preloaded with protein X degraded their DNA more slowly. We propose that protein X could protect DNA against degradation by binding to singlestranded regions, thereby inhibiting nuclease action.
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Boyle, J.V., Cook, T.M., Goss, W.A.: Mechanism of action of nalidixic acid on Escherichia coli. VI. Cell-free studies. J. Bacteriol 97, 230–236 (1969)
Cook, T.M., Deitz, W.M., Goss, W.A.: Mechanism of action of nalidixic acid on Escherichia coli. IV. Effects on the stability of cellular constituents. J. Bacteriol. 91, 774–779 (1966)
Dressler, D.: The recent excitement in the DNA growing point problem. Annu. Rev. Microbiol. 29, 525–559 (1975)
Fietta, A.M., Silvestri, L.G.: Mechanism of action of rifamazine, a member of a new class of (dimetric) rifamycins. Eur. J. Biochem. 52, 391–400 (1975)
Gefter, M.L.: DNA replication. Ann. Rev. Biochem. 44, 45–78 (1975)
Gudas, L.J.: The induction of protein X in DNA repair and cell division mutants of Escherichia coli. J. Mol. Biol. 104, 567–587 (1976)
Gudas, L.J., Mount, D.W.: Identification of the recA (tif) gene product of Escherichia coli. Proc. Natl. Acad. Sci. USA. 74, 5280–5284 (1977)
Gudas, L.J., Mount, D.W.: Manuscript in preparation (on lexA mutants) (1978)
Gudas, L.J., Pardee, A.B.: Model for regulation of Escherichia coli DNA repair functions. Proc. Natl. Acad. Sci. USA. 72, 2330–2334 (1975)
Gudas, L.J., Pardee, A.B.: DNA synthesis inhibition and the induction of protein X in Escherichia coli. J. Mol. Biol. 101, 459–477 (1976)
Haidle, C.W.: Fragmentation of deoxyribonucleic acid by bleomycin. Mol. Pharmacol. 7, 545–552 (1971)
Hall, J.D., Howard-Flanders, P.: Temperature sensitive recA: deoxyribonucleic acid metabolism after ultraviolet irradiation. J. Bacteriol. 121, 892–900 (1975)
Hill, W.E., Fangman, W.L.: Single-strand breaks in deoxyribonucleic acid and viability loss during deoxyribonucleic acid synthesis inhibition in Escherichia coli. J. Bacteriol. 116, 1329–1335 (1973)
Howard-Flanders, P., Theriot, L.: Mutants of Escherichia coli K12 deficient in DNA repair and in genetic recombination. Genetics 53, 1137–1150 (1966)
Inouye, M.: Pleiotypic effect of the recA gene of Escherichia coli: uncoupling of cell division from deoxyribonucleic acid replication. J. Bacteriol. 106, 539–542 (1971)
Inouye, M., Pardee, A.B.: Changes of membrane proteins and their relation of deoxyribonucleic acid synthesis and cell division of Escherichia coli. J. Biol. Chem. 245, 5813–5819 (1970)
Laemmli, U.K.: Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680–685 (1970)
Leive, L.: Actinomycin sensitivity in Escherichia coli produced by EDTA. Biochem. Biophys. Res. Commun. 18, 13–17 (1965)
Little, J.W., Hanawalt, P.C.: Induction of protein X in Escherichia coli. Mol. Gen. Genet. 150, 237–248 (1977)
Little, J.W., Kleid, D.G.: E. coli protein X is the recA gene product. J. Biol. Chem. 252, 6251–6252 (1977)
Luria, S.E., Adams, J.N., Ting, R.C.: Transduction of lactose-utilizing ability among strains of E. coli and S. dysenteriae and the properties of the transducing phage particles. Virology 12, 348–390 (1960)
Mackay, V., Linn, S.: Selective inhibition of the DNAse activity of the recBC enzyme by the DNA binding protein from Escherichia coli. J. Biol. Chem. 251, 3716–3719 (1976)
Marsden, H.S., Pollard, E.C., Ginoza, W., Randall, E.P.: Involvement of recA and exr genes in the in vivo inhibition of the recBC nuclease. J. Bacteriol. 118, 465–470 (1974)
McEntee, K.: Protein X is the product of the recA gene of Escherichia coli. Proc. Natl. Acad. Sci. USA. 74, 5275–5279 (1977)
Mount, D.: 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 (1977)
Oishi, M.: An ATP-dependent deoxyribonuclease from Escherichia coli with a possible role in genetic recombination. Proc. Natl. Acad. Sci. USA. 64, 1292–1299 (1969)
Ramareddy, G., Reiter, H.: Specific loss of newly replicated deoxyribonucleic acid in nalidixic acid treated Bacillus subtilis 168. J. Bacteriol. 100, 724–729 (1969)
Roberts, J.W., Roberts, C.W.: Proteolytic cleavage of bacteriophage lambda repressor in induction. Proc. Natl. Acad. Sci. USA. 72, 147–151 (1975)
Ross, S.L., Moses, R.E.: Effect of bleomycin on deoxyribonucleic acid synthesis in toluene-treated Escherichia coli cells. Antimicrob. Agents Chemother. 9, 239–246 (1976)
Satta, G., Pardee, A.B.: Inhibition of Escherichia coli division by protein X. J. Bacteriol. 133, 1492–1500 (1978)
Sedgwick, S.G.: Ultraviolet inducible protein associated with error prone repair in E. coli B. Nature 255, 349–350 (1975)
Siccardi, A.G., Shapiro, B.M., Hirota, X., Jacob, F.: On the process of cellular division in Escherichia coli. IV. Altered protein composition and turnover of the membranes of thermosensitive mutants defective in chromosomal replication. J. Mol. Biol. 56, 475–490 (1971)
Smith, C.L., Oishi, M.: Early events and mechanisms in the induction of bacterial S.O.S. functions: analysis of the phage repressor inactivation process in vivo. Proc. Natl. Acad. Sci. USA. 75, 1657–1661 (1978)
Sussman, R., Ben Zeev, H.: Proposed mechanism of bacteriophage lambda induction: acquisition of binding sites for lambda repressor by DNA of the host. Proc. Natl. Acad. Sci. USA. 72, 1973–1976 (1975)
Suzuki, H., Nagai, R., Yamaki, H., Tanaka, N., Umezawa, H.: Mechanism of action of bleomycin: scission of DNA strands in vivo and in vitro. J. Antibiot. (Tokyo) 22, 446–448 (1969)
Volkert, M.R., George, D.L., Witkin, E.M.: Partial suppression of the lexA phenotype by mutations (rnm) which restore ultraviolet resistance but not ultraviolet mutability to Escherichia coli B/r uvrA lexA. Mutat. Res. 36, 17–28 (1976)
Witkin, E.M.: Ultraviolet mutagenesis and inducible DNA repair in Escherichia coli. Bacteriol. Rev. 40, 869–907 (1976)
Wozny, M.E., Carnevale, H.N., Jones, E.E.: Alteration of regulation of arginine biosynthesis in Escherichia coli W by mutation to rifampicin resistance. Biochim. Biophys. Acta 383, 106–116 (1975)
Yasbin, R.E.: DNA repair in Bacillus subtilis. Mol. Gen. Genet. 153, 211–218; 219–225 (1977)
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Communicated by A.J. Clark
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Satta, G., Gudas, L.J. & Pardee, A.B. Degradation of Escherichia coli DNA: Evidence for limitation in vivo by protein X, the recA gene product. Molec. Gen. Genet. 168, 69–80 (1979). https://doi.org/10.1007/BF00267935
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DOI: https://doi.org/10.1007/BF00267935