Summary
Recombination in vivo was studied in recA - heterozygous lacZ merodiploids by performing β-galactosidase assays after infection with λprecA +. Recombination as measured by β-galactosidase production was a linear function of λpecA + multiplicity of infection (MOI) when the strain contained a deletion of the chromosomal recA gene. However, when the strain carried a recA1 missense allele, a higher λprecA + MOI was required to obtain levels of recombination comparable to the Δ(recA) strain, and the slope of the dose-response curve increased to approximately two. It is proposed that negative complementation occurs in mixed tetramers of wild-type and missense recA polypeptides, and that in vivo recombination is a property of a multimeric form of recA protein.
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References
Bachmann B (1972) Pedigrees of some mutant strains of Escherichia coli K-12. Bacteriol Rev 36:525–557
Birge EA, Low KB (1974) Detection of transcribable recombination products following conjugation in Rec+, RecB- and RecC- strains of Escherichia coli K-12. J Mol Biol 83:447–457
Charette MF, Henderson GW, Kezdy FJ, Markovitz A (1982) Molecular mechanism for dominance of a mutant allele of an ATP-dependent protease. J Mol Biol 162:503–510
Cox M, 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
DasGupta C, Shibata T, Cunningham R, Radding CM (1980) The topology of homologous pairing promoted by recA protein. Cell 22:437–446
DasGupta C, Wu A, Kahn R, Cunningham R, Radding CM (1981) Concerted strand exchange and formation of Holliday structures by E. coli recA protein. Cell 25:507–516
Little JW, Mount D (1982) The SOS regulatory system of Escherichia coli. Cell 29:11–22
Lloyd RG, Low B (1976) Some genetic consequences of changes in the level of recA gene function in Escherichia coli K-12. Genetics 84:675–695
Low B (1973) Rapid mapping of conditional and auxotrophic mutations in Escherichia coli K-12. J Bacteriol 113:798–812
McEntee K, Weinstock G, 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
McEntee K, Weinstock G, Lehman IR (1981) Binding of the recA protein in Escherichia coli to single- and double-stranded DNA. J Biol Chem 256:8835–8844
Miller JH (1972) Experiments in Molecular Genetics. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y.
Mount D (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
Müller-Hill B, Crapo L, Gilbert W (1968) Mutants that make more lac repressor. Proc Natl Acad Sci USA 59:1259–1264
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
Porter RD, McLaughlin T, Low B (1978) Transduction versus “Conjuduction”: evidence for multiple roles for exonuclease V in genetic recombination in Escherichia coli. Cold Spring Harbor Symp Quant Biol 43:1043–1047
Porter RD, Lark M, Low KB (1981) Specialized transduction with λplac5: dependence on recA and on configuration of lac and attλ. J Virology 38:497–503
Ogawa T, Wabiko 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
Radding CM, Shibata T, DasGupta C, Cunningham R, Osber L (1980) Kinetics and topology of homologous pairing promoted by Escherichia coli recA protein. Cold Spring Harbor Symp Quant Biol 45:385–390
Radding CM (1982) Homologous pairing and strand exchange in genetic recombination. Annu Rev Genet 16:405–437
Weinstock G, 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
West S, Cassuto E, Howard-Flanders P (1981a) recA protein promotes homologous-pairing and strand-exchange reactions between duplex DNA molecules. Proc Natl Acad Sci USA 78:2100–2104
West S, Cassuto E, Howard-Flanders P (1981b) Heteroduplex formation by recA protein: polarity of strand exchanges. Proc Natl Acad Sci USA 78:6149–6153
Willis DK, Uhlin BG, Amini KS, Clark AJ (1981) Physical mapping of the srl recA region of Escherichia coli: analysis of Tn10 generated insertions and deletions. Molec Gen Genet 183:497–504
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Yancey, S.D., Porter, R.D. Negative complementation of recA protein by recA1 polypeptide: in vivo recombination requires a multimeric form of recA protein. Molec. Gen. Genet. 193, 53–57 (1984). https://doi.org/10.1007/BF00327413
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DOI: https://doi.org/10.1007/BF00327413