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. 1981 Jun;78(6):3433–3437. doi: 10.1073/pnas.78.6.3433

recA protein of Escherichia coli promotes branch migration, a kinetically distinct phase of DNA strand exchange.

M M Cox, I R Lehman
PMCID: PMC319582  PMID: 7022448

Abstract

The recA protein of Escherichia coli promotes the complete exchange of strands between full-length linear duplex and single-stranded circular DNA molecules of bacteriophage phi X-174, converting more than 50% of the single-stranded DNA into heteroduplex replicative form II-like structures. Kinetically, the reaction can be divided into two phases, formation of short heteroduplex regions (D loops) and extension of the D loops via branch migration. recA protein participates directly in both phases. D loops are formed efficiently in the presence of ATP or the nonhydrolyzable ATP analog adenosine 5'-[gamma-thio]triphosphate, whereas D-loop extension requires continuous ATP hydrolysis. Complete strand exchange requires a stoichiometric amount of recA protein and is strongly stimulated by the single-stranded-DNA-binding protein of E. coli.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Beattie K. L., Wiegand R. C., Radding C. M. Uptake of homologous single-stranded fragments by superhelical DNA. II. Characterization of the reaction. J Mol Biol. 1977 Nov;116(4):783–803. doi: 10.1016/0022-2836(77)90271-6. [DOI] [PubMed] [Google Scholar]
  2. Broker T. R., Lehman I. R. Branched DNA molecules: intermediates in T4 recombination. J Mol Biol. 1971 Aug 28;60(1):131–149. doi: 10.1016/0022-2836(71)90453-0. [DOI] [PubMed] [Google Scholar]
  3. Clark A. J. Recombination deficient mutants of E. coli and other bacteria. Annu Rev Genet. 1973;7:67–86. doi: 10.1146/annurev.ge.07.120173.000435. [DOI] [PubMed] [Google Scholar]
  4. Cunningham R. P., DasGupta C., Shibata T., Radding C. M. Homologous pairing in genetic recombination: recA protein makes joint molecules of gapped circular DNA and closed circular DNA. Cell. 1980 May;20(1):223–235. doi: 10.1016/0092-8674(80)90250-0. [DOI] [PubMed] [Google Scholar]
  5. DasGupta C., Shibata T., Cunningham R. P., Radding C. M. The topology of homologous pairing promoted by RecA protein. Cell. 1980 Nov;22(2 Pt 2):437–446. doi: 10.1016/0092-8674(80)90354-2. [DOI] [PubMed] [Google Scholar]
  6. Holloman W. K., Radding C. M. Recombination promoted by superhelical DNA and the recA gene of Escherichia coli. Proc Natl Acad Sci U S A. 1976 Nov;73(11):3910–3914. doi: 10.1073/pnas.73.11.3910. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Horii T., Ogawa T., Ogawa H. Organization of the recA gene of Escherichia coli. Proc Natl Acad Sci U S A. 1980 Jan;77(1):313–317. doi: 10.1073/pnas.77.1.313. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Hotchkiss R. D. Models of genetic recombination. Annu Rev Microbiol. 1974;28(0):445–468. doi: 10.1146/annurev.mi.28.100174.002305. [DOI] [PubMed] [Google Scholar]
  9. Lee C. S., Davis R. W., Davidson N. A physical study by electron microscopy of the terminally reptitious, circularly permuted DNA from the coliphage particles of Escherichia coli 15. J Mol Biol. 1970 Feb 28;48(1):1–22. doi: 10.1016/0022-2836(70)90215-9. [DOI] [PubMed] [Google Scholar]
  10. McEntee K., Hesse J. E., Epstein W. Identification and radiochemical purification of the recA protein of Escherichia coli K-12. Proc Natl Acad Sci U S A. 1976 Nov;73(11):3979–3983. doi: 10.1073/pnas.73.11.3979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. McEntee K., Weinstock G. M., Lehman I. R. Initiation of general recombination catalyzed in vitro by the recA protein of Escherichia coli. Proc Natl Acad Sci U S A. 1979 Jun;76(6):2615–2619. doi: 10.1073/pnas.76.6.2615. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. McEntee K., Weinstock G. M., Lehman I. R. recA protein-catalyzed strand assimilation: stimulation by Escherichia coli single-stranded DNA-binding protein. Proc Natl Acad Sci U S A. 1980 Feb;77(2):857–861. doi: 10.1073/pnas.77.2.857. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Ogawa T., Wabiko H., Tsurimoto T., Horii T., Masukata H., Ogawa H. Characteristics of purified recA protein and the regulation of its synthesis in vivo. Cold Spring Harb Symp Quant Biol. 1979;43(Pt 2):909–915. doi: 10.1101/sqb.1979.043.01.099. [DOI] [PubMed] [Google Scholar]
  14. Radding C. M., Beattie K. L., Holloman W. K., Wiegand R. C. Uptake of homologous single-stranded fragments by superhelical DNA. IV. Branch migration. J Mol Biol. 1977 Nov;116(4):825–839. doi: 10.1016/0022-2836(77)90273-x. [DOI] [PubMed] [Google Scholar]
  15. Radding C. M. Genetic recombination: strand transfer and mismatch repair. Annu Rev Biochem. 1978;47:847–880. doi: 10.1146/annurev.bi.47.070178.004215. [DOI] [PubMed] [Google Scholar]
  16. Roberts J. W., Roberts C. W., Craig N. L., Phizicky E. M. Activity of the Escherichia coli recA-gene product. Cold Spring Harb Symp Quant Biol. 1979;43(Pt 2):917–920. doi: 10.1101/sqb.1979.043.01.100. [DOI] [PubMed] [Google Scholar]
  17. Sancar A., Stachelek C., Konigsberg W., Rupp W. D. Sequences of the recA gene and protein. Proc Natl Acad Sci U S A. 1980 May;77(5):2611–2615. doi: 10.1073/pnas.77.5.2611. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Shibata T., Cunningham R. P., DasGupta C., Radding C. M. Homologous pairing in genetic recombination: complexes of recA protein and DNA. Proc Natl Acad Sci U S A. 1979 Oct;76(10):5100–5104. doi: 10.1073/pnas.76.10.5100. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Shibata T., DasGupta C., Cunningham R. P., Radding C. M. Homologous pairing in genetic recombination: formation of D loops by combined action of recA protein and a helix-destabilizing protein. Proc Natl Acad Sci U S A. 1980 May;77(5):2606–2610. doi: 10.1073/pnas.77.5.2606. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Shibata T., DasGupta C., Cunningham R. P., Radding C. M. Purified Escherichia coli recA protein catalyzes homologous pairing of superhelical DNA and single-stranded fragments. Proc Natl Acad Sci U S A. 1979 Apr;76(4):1638–1642. doi: 10.1073/pnas.76.4.1638. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Simpson P. A., Spudich J. A. ATP-driven steady-state exchange of monomeric and filamentous actin from Dictyostelium discoideum. Proc Natl Acad Sci U S A. 1980 Aug;77(8):4610–4613. doi: 10.1073/pnas.77.8.4610. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Sodergren E. J., Fox M. S. Effects of DNA sequence non-homology on formation of bacteriophage lambda recombinants. J Mol Biol. 1979 Jun 5;130(4):357–377. doi: 10.1016/0022-2836(79)90428-5. [DOI] [PubMed] [Google Scholar]
  23. Thompson B. J., Camien M. N., Warner R. C. Kinetics of branch migration in double-stranded DNA. Proc Natl Acad Sci U S A. 1976 Jul;73(7):2299–2303. doi: 10.1073/pnas.73.7.2299. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Weinstock G. M., McEntee K., Lehman I. R. ATP-dependent renaturation of DNA catalyzed by the recA protein of Escherichia coli. Proc Natl Acad Sci U S A. 1979 Jan;76(1):126–130. doi: 10.1073/pnas.76.1.126. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. West S. C., Cassuto E., Howard-Flanders P. recA protein promotes homologous-pairing and strand-exchange reactions between duplex DNA molecules. Proc Natl Acad Sci U S A. 1981 Apr;78(4):2100–2104. doi: 10.1073/pnas.78.4.2100. [DOI] [PMC free article] [PubMed] [Google Scholar]

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