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. 1981 Aug 25;9(16):4201–4210. doi: 10.1093/nar/9.16.4201

Genetic recombination: recA protein promotes homologous pairing between duplex DNA molecules without strand unwinding.

E Cassuto, S C West, J Podell, P Howard-Flanders
PMCID: PMC1058476  PMID: 7029464

Abstract

The recA protein of Escherichia coli promotes pairing in vitro between covalent circular duplex DNA and homologous circular duplex DNA containing a single stranded region. We have used a filter binding assay to investigate the frequency of homologous pairing between gapped and intact duplex DNA when unwinding of the free 3' and 5' ends of the gapped molecules was blocked. In order to obtain DNA without free ends, the gapped DNA was treated with trimethylpsoralen and 360 nm light so as to introduce about 6 crosslinks per DNA molecule and the double stranded regions on either side of the gaps were then digested up to the first crosslinks with exonuclease III and lambda exonuclease. This treatment did not diminish the frequency of homologous pairing, an observation which is difficult to reconcile with models for recombination requiring strand unwinding before pairing.

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Selected References

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

  1. Cassuto E., Gross N., Bardwell E., Howard-Flanders P. Genetic effects of photoadducts and photocross-links in the DNA of phage lambda exposed to 360 nm light and tri-methylpsoralen or khellin. Biochim Biophys Acta. 1977 Apr 19;475(4):589–600. doi: 10.1016/0005-2787(77)90319-7. [DOI] [PubMed] [Google Scholar]
  2. Cassuto E., Mursalim J., Howard-Flanders P. Homology-dependent cutting in trans of DNA in extracts of Escherichia coli: an approach to the enzymology of genetic recombination. Proc Natl Acad Sci U S A. 1978 Feb;75(2):620–624. doi: 10.1073/pnas.75.2.620. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Cassuto E., West S. C., Mursalim J., Conlon S., Howard-Flanders P. Initiation of genetic recombination: homologous pairing between duplex DNA molecules promoted by recA protein. Proc Natl Acad Sci U S A. 1980 Jul;77(7):3962–3966. doi: 10.1073/pnas.77.7.3962. [DOI] [PMC free article] [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. 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]
  6. McGavin S. Models of specifically paired like (homologous) nucleic acid structures. J Mol Biol. 1971 Jan 28;55(2):293–298. doi: 10.1016/0022-2836(71)90201-4. [DOI] [PubMed] [Google Scholar]
  7. Meselson M. S., Radding C. M. A general model for genetic recombination. Proc Natl Acad Sci U S A. 1975 Jan;72(1):358–361. doi: 10.1073/pnas.72.1.358. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Morgan A. R., Wells R. D. Specificity of the three-stranded complex formation between double-stranded DNA and single-stranded RNA containing repeating nucleotide sequences. J Mol Biol. 1968 Oct 14;37(1):63–80. doi: 10.1016/0022-2836(68)90073-9. [DOI] [PubMed] [Google Scholar]
  9. Rupp W. D., Wilde C. E., 3rd, Reno D. L., Howard-Flanders P. Exchanges between DNA strands in ultraviolet-irradiated Escherichia coli. J Mol Biol. 1971 Oct 14;61(1):25–44. doi: 10.1016/0022-2836(71)90204-x. [DOI] [PubMed] [Google Scholar]
  10. 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]
  11. West S. C., Cassuto E., Howard-Flanders P. Homologous pairing can occur before DNA strand separation in general genetic recombination. Nature. 1981 Mar 5;290(5801):29–33. doi: 10.1038/290029a0. [DOI] [PubMed] [Google Scholar]
  12. 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]
  13. West S. C., Cassuto E., Mursalim J., Howard-Flanders P. Recognition of duplex DNA containing single-stranded regions by recA protein. Proc Natl Acad Sci U S A. 1980 May;77(5):2569–2573. doi: 10.1073/pnas.77.5.2569. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Wilson J. H. Nick-free formation of reciprocal heteroduplexes: a simple solution to the topological problem. Proc Natl Acad Sci U S A. 1979 Aug;76(8):3641–3645. doi: 10.1073/pnas.76.8.3641. [DOI] [PMC free article] [PubMed] [Google Scholar]

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