Summary
We have transduced the mutant allele ssb-1, which encodes a temperature-sensitive single-strand DNA binding protein (SSB), into several Escherichia coli strains, and have examined colony-forming ability, DNA replication, sensitivity to ultraviolet light (UV) and UV-induced mutability at the nonpermissive temperature. We have found: 1) that the degree of ssb-1-mediated temperature-sensitivity of colony-forming ability and of DNA replication is strain-dependent, resulting in plating efficiencies at 42° C (relative to 30° C) ranging from 100% to 0.002%; 2) that complete suppression of the temperature-sensitivity caused by ssb-1 occurs only on nutrient agar, and not in any other medium tested; 3) that strains in which ssb-1-mediated temperature-sensitivity is completely suppressed show moderate UV sensitivity and normal UV mutability at 30° C, but much more extreme UV sensitivity and drastically reduced UV mutability at 42° C; and 4) that defects in excision repair or in other Uvr+-dependent processes are not responsible for most of the UV sensitivity promoted by ssb-1. We discuss our results in relation to the known properties of SSB and its possible role in the induction of DNA damage-inducible (SOS) functions.
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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
Bockrath RC, Palmer JE (1977) Differential repair of premutational UV-lesions at tRNA genes in E. coli. Mol Gen Genet 156:133–140
Brent R, Ptashe M (1981) Mechanism of action of the lexA gene product. Proc Natl Acad Sci USA (in press)
Clarke CH, Hill RF (1972) Mutation frequency decline for streptomycin-resistant mutations induced by ultraviolet light in Escherichia coli B/r. Mutat Res 14:247–249
Craig NL, Roberts JW (1980) E. coli recA protein-directed cleavage of phage λ repressor requires polynucleotide. Nature 283:26–30
Emmierson PT, West SC (1977) Identification of protein X of Escherichia coli as the recA +/tif + gene product. Mol Gen Genet 155:77–85
George DL, Witkin EM (1974) Slow excision repair in an mfd mutant of Escherichia coli B/r. Mol Gen Genet 133:283–291
Glassberg J, Meyer RR, Kornberg A (1979) Mutant single-strand binding protein of Escherichia coli: genetic and physiological characterization. J Bacteriol 140:14–19
Gudas LJ, Mount DW (1977) Identification of the recA (tif) gene product of Escherichia coli. Proc Natl Acad Sci USA 74:5280–5284
Hill RF (1965) Ultraviolet-induced lethality and reversion to prototrophy in Escherichia coli strains with normal and reduced dark repair ability. Photochem Photobiol 4:563–568
Howard-Flanders P, Boyce RP (1966) DNA repair and genetic recombination: studies on mutants of Escherichia coli defective in these processes. Radiat Res 6:(suppl) 156–184
Johnson BF (1977a) Fine structure mapping and properties of mutations suppressing the lon mutation in Escherichia coli K-12 and B strains. Genet Res, Camb 30:273–286
Johnson BF (1977b) Genetic mapping of the lexC-113 mutation. Mol Gen Genet 157:91–97
Kondo S, Kato T (1966) Action spectra for photoreactivation of killing and mutation to prototrophy in UV-sensitive strain of Escherichia coli possessing and lacking photoreactivating enzyme. Photochem Photobiol 5:827–837
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
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 (in press)
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
Meyer RR, Glassberg J, Kornberg A (1979) An Escherichia coli mutant defective in single-strand binding protein is defective in DNA replication. Proc Natl Acad Sci USA 76:1702–1705
Meyer RR, Glassberg J, Scott JV, Kornberg A (1980) A temperature-sensitive single-stranded DNA-binding protein from Escherichia coli. J Biol Chem 255:2897–2901
Miller JH (1972) Experiments in molecular genetics. Cold Spring Harbor Laboratory, New York
Molineux IJ, Friedman S, Gefter ML (1974) Purification and properties of the Escherichia coli deoxyribonucleic acid-unwinding protein. Effects on deoxyribonucleic acid synthesis in vitro. J Biol Chem 249:6090–6098
Mosig G, Breschkin AM (1975) Genetic evidence for an additional function of phage T4 gene 32 protein: interaction with ligase. Proc Natl Acad Sci USA 72:1226–1230
Mount DW, Low KB, Edmiston SH (1972) Dominant mutations (lex) in Escherichia coli K-12 which affect radiation sensitivity and frequency of ultraviolet light-induced mutations. J Bacteriol 112:886–893
Roberts JW, Roberts CW, Craig NL (1978) Escherichia coli recA gene inactivates λ repressor. Proc Natl Acad Sci USA 75:4714–4718
Rupp WD, Wilde CE III, Reno DL, Howard-Flanders P (1971) Exchanges between DNA strands in ultraviolet-irradiated Escherichia coli. J Mol Biol 61:25–44
Sedgwick SG, Levine A, Bailone A (1978) Induction of recA +-protein synthesis in Escherichia coli. Mol Gen Genet 160:267–276
Shibata T, DasGupta C, Cunningham RP, Radding CM (1980) Homologous pairing in genetic recombination: formation of D loops by combined action of recA protein and a helix-destabilizing protein. Proc Natl Acad Sci USA 77:2606–2610
Sigal N, Delius H, Kornberg T, Gefter ML, Alberts B (1972) A DNA-unwinding protein isolated from Escherichia coli: its interaction with DNA and with DNA polymerases. Proc Natl Acad Sci USA 69:3537–3541
Sussman R, Resnick J, Calame K, Baluch J (1978) Interaction of bacteriophage lambda repressor with non-operator DNA containing single strand gaps. Proc Natl Acad Sci USA 75:5817–5821
Vales LD, Chase JW, Murphy JB (1980) Effect of the ssbAl and lexC113 mutations on lambda prophage induction, phage growth and cell survival. J Bacteriol 143:887–896
Vogel HJ, Bonner DM (1956) Acetylornithinase of Escherichia coli: partial purification and some properties. J Biol Chem 218:97–106
Volkert M, 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 uvrA lexA. Mutat Res 36:17–28
Weiner JH, Bertsch LL, Kornberg A (1975) The deoxyribonucleic acid unwinding protein of Escherichia coli. Properties and functions in replication. J Biol Chem 250:1972–1980
Whittier RF, Chase JW (1980) A filter assay specific to Eco helixdestabilizing-protein I in crude extracts. Anal Biochem 106:99–108
Whittier RF, Chase JW (1981) DNA repair in E. coli strains deficient in single-strand DNA binding protein. Mol Gen Genet 183:341–347
Witkin EM (1947) Genetics of resistance to radiation in Escherichia coli. Genetics 32:221–248
Witkin EM (1964) Photoreversal and “dark repair” of mutations to prototrophy induced by ultraviolet light in photoreactivable and non-photoreactivable strains of Escherichia coli. Mutat Res 1:22–36
Witkin EM (1964) Radiation-induced mutations and their repair. Science 152:1345–1353
Witkin EM (1967) Mutation proof and mutation prone modes of survival in derivatives of Escherichia coli B differing in sensitivity to ultraviolet light. Brookhaven Symp Biol 20:17–55
Witkin EM (1969) Ultraviolet-induced mutation and DNA repair. Annu Rev Microbiol 23:487–514
Witkin EM (1974) Thermal enhancement of ultraviolet mutability in a tif-1 uvrA derivative of Escherichia coli B/r: evidence that ultraviolet mutagenesis depends upon an inducible function. Proc Natl Acad Sci USA 71:1930–1934
Witkin EM (1975) Persistence and decay of thermoinducible errorprone repair activity in nonfilamentous derivatives of tif-1 Escherichia coli B/r: the timing of some critical events in ultraviolet mutagenesis. Mol Gen Genet 142:87–103
Witkin EM (1976) Ultraviolet mutagenesis and inducible deoxyribonucleic acid repair in Escherichia coli. Bacteriol Rev 40:869–907
Witkin EM, Sicurella NA, Bennet GM (1963) Photoreversibility of induced mutations in a nonphotoreactivable strain of Escherichia coli. Proc Natl Acad Sci USA 50:1055–1058
Witkin EM, Wermundsen IE (1977) Induction of lambda prophage and of mutations to streptomycin resistance in separate small fractions of a lysogenic derivative of Escherichia coli B/r by very low doses of ultraviolet light. Mol Gen Genet 156:35–39
Wood WJ Jr, Bernstein H (1977) Suppressors of gene 32 am mutants that specifically overproduce P32 (unwinding protein) in bacteriophage T4. J Virol 21:619–625
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Communicated by B.A. Bridges
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Lieberman, H.B., Witkin, E.M. 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. Molec. Gen. Genet. 183, 348–355 (1981). https://doi.org/10.1007/BF00270639
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DOI: https://doi.org/10.1007/BF00270639