Summary
The sdrA102 mutation confers upon cells the ability to replicate DNA in the absence of protein synthesis. This mutation was combined with the recA200 mutation, which renders the recA protein thermolabile, and had little effect on normal replication. However, the sdrA102 recA200 double mutant exhibited temperature-sensitive stable DNA replication: it replicated DNA continuously in the presence of chloramphenicol at 30°C, whereas at 42°C DNA replication ceased after the DNA content increased only 40–45%. Suppressor mutants (rin; recA-independent) capable of stable DNA replication at 42°C were isolated from the double mutant. The suppressor mutant retained all other recA − characteristics, i.e., deficient general recombination, severe UV-sensitivity, and incapability of prophage induction in lysogens. This indicates that the rin mutation specifically suppresses the recA + dependency of stable DNA replication. It is suggested that the recA + protein stabilizes a specific structure, similar to an intermediate in recombination, which may function in the initiation of stable DNA replication.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Benbow RM, Hutchinson CA, Fabricant JD, Sinsheimer RL (1971) Genetic map of bacteriophage ϕX174. J Virol 7:549–558
Caro L, Berg CM (1968) Chromosome replication in some strains of E. coli K-12. Cold Spring Harbor Symp Quant Biol 33:559–573
Clark AJ, Chamberlin M, Boyce RP, Howard-Flanders P (1966) Abnormal metabolic response to ultraviolet light of a recombination-deficient mutant of Escherichia coli K-12. J Mol Biol 19:442–454
Craig NL, Roberts JW (1980) E. coli recA protein-directed cleavage of phage λ repressor requires polynucleotide. Nature (London) 283:26–29
Csonka LN, Clark AJ (1979) Deletions generated by the transposon Tn10 in the srl recA region of the Escherichia coli K-12 chromosome. Genetics 93:321–343
Csonka LN, Clark AJ (1980) Construction of an Hfr strain useful for transferring recA mutations between Escherichia coli strains. J Bacteriol 143:529–530
Devoret R, Blanco M (1970) Mutants of Escherichia coli K-12 (λ) non-inducible by thymine deprivation. 1. Method of isolation and classes of mutants obtained. Mol Gen Genet 107:272–280
Froehlich B, Epstein W (1981) Escherichia coli mutants in which transcription is dependent on recA function. J Bacteriol 147:1117–1120
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
Kenyon CJ, Walker GC (1981) Expression of the E. coli uvrA gene is inducible. Nature (London) 289:808–810
Kilbane JJ, Malamy MH (1980) F factor mobilization of nonconjugative chimeric plasmids in Escherichia coli: general mechanism and a role for site-specific recA-independent recombination at oriV. J Mol Biol 143:73–93
Kogoma T (1978) A novel Escherichia coli mutant capable of DNA replication in the absence of protein synthesis. J Mol Biol 121:55–69
Kogoma T, Lark KG (1970) DNA replication in Escherichia coli: replication in absence of protein synthesis after replication inhibition. J Mol Biol 52:143–164
Kogoma T, Lark KG (1975) Characterization of the replication of Escherichia coli DNA in the absence of protein synthesis: stable DNA replication. J Mol Biol 94:243–256
Kogoma T, Torrey TA, Connaughton MJ (1979) Induction of UV-resistant DNA replication in Escherichia coli: induced stable DNA replication as an SOS function. Mol Gen Genet 176:1–9
Kogoma T, Torrey TA, Subia NL, Pickett GG An alternative DNA initiation pathway in E. coli. In: Ray DS (ed) The Initiation of DNA Replication (Academic Press, New York), pp 361–374
Lark KG, Lark C (1978) recA-dependent DNA replication in the absence of protein synthesis: characteristics of a dominant lethal replication mutation, dnaT and requirement for recA + function. Cold Spring Harbor Symp Quant Biol 43:537–549
LaVerne LS, Ray DS (1980) Site-specific integration of an F′ lacpro factor in the region of the replication origin (oriC) of E. coli. Mol Gen Genet 179:437–446
Little JW, Edmiston SH, Pacelli LZ, Mount DW (1980) Cleavage of the Escherichia coli lexA protein by the lexA 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 78:4199–4203
Lloyd RG, Low B, Godson GN, Birge EA (1974) Isolation and characterization of an Escherichia coli K-12 mutant with a temperature-sensitive RecA− phenotype. J Bacteriol 120:407–415
Lloyd RG, Low B (1976) Some genetic consequences of changes in the levels of recA gene function in Escherichia coli K-12. Genetics 84:675–695
Masters M, Andresdottir V, Wolf-Watz H (1978) Plasmids carrying oriC can integrate at or near the chromosome origin of Escherichia coli in the absence of a functional recA product. Cold Spring Harbor Symp Quant Biol 43:1069–1072
McEntee K, Weinstock GM, Lehman IR (1979) Initiation of general recombination catalyzed in vitro by the recA protein of Escherichia coli. Proc Natl Acad Sci USA 76:2615–2619
Miller JH (1972) Experiments in molecular genetics. Cold Spring Harbor Laboratory: Cold Spring Harbor, New York
Morand P, Blanco M, Devoret R (1977) Characterization of lexB mutations in Escherichia coli K-12. J Bacteriol 131:572–582
Mosig G, Luder A, Rowen L, Macdonald P, Bock S (1981) On the role of recombination and topoisomerase in primary and secondary initiation of T4 DNA replication. In: Ray DS (ed) The Initiation of DNA Replication. (Academic Press, New York), pp 211–295
Ogawa T, Wabico H, Tsurimoto T, Horii T, Masukata H, Ogawa H (1978) Characteristics of purified recA protein and the regulation of its synthesis in vivo. Cold Spring Harbor Symp Quant Biol 43:909–915
Roberts JW, Roberts CW, Craig NL, Phizicky EM (1978a) Activity of the Escherichia coli recA gene product. Cold Spring Harbor Symp Quant Biol 43:917–920
Roberts JW, Roberts CW, Craig NL (1978b) Escherichia coli recA gene product inactivates phage lamda repressor. Proc Natl Acad Sci USA 75:4714–4718
Roberts JW, Roberts CW (1981) Two mutations that alter the regulatory activity of E. coli recA protein. Nature (London) 290:422–424
Shibata T, DasGupta C, Cunningham RP, Radding CM (1979) Purified E. coli recA protein catalyzes homologous pairing of superhelical DNA and single-stranded fragments. Proc Natl Acad Sci USA 76:1638–1642
Stacey KA, Simson E (1965) Improved method for the isolation of thymine-requiring mutants of Escherichia coli. J Bacteriol 90:554–555
Willetts NS, Clark AJ (1969) Characteristics of some multiply recombination-deficient strains of E. coli. J Bacteriol 100:231–239
Witkin EM (1976) Ultraviolet mutagenesis and inducible DNA repair in Escherichia coli. Bacteriol Rev 40:869–907
Author information
Authors and Affiliations
Additional information
Communicated by G. O'Donovan
Rights and permissions
About this article
Cite this article
Torrey, T.A., Kogoma, T. Suppressor mutations (rin) that specifically suppress the recA + dependence of stable DNA replication in Escherichia coli K-12. Molec Gen Genet 187, 225–230 (1982). https://doi.org/10.1007/BF00331121
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00331121