Homologous pairing and topological linkage of DNA molecules by combined action of E. coli recA protein and topoisomerase I

Cunningham, Richard P. ; Wu, Anna M. ; Shibata, Takehiko ; Dasgupta, Chanchal ; Radding, Charles M. (1981) Homologous pairing and topological linkage of DNA molecules by combined action of E. coli recA protein and topoisomerase I Cell, 24 (1). pp. 213-223. ISSN 0092-8674

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Official URL: http://www.sciencedirect.com/science/article/pii/0...

Related URL: http://dx.doi.org/10.1016/0092-8674(81)90517-1


E. coil RecA protein and topolsomerase I, acting on superhelical DNA and circular single strands in the presence of ATP and Mg2+, topologically link single-stranded molecules to one another, and single-stranded molecules to duplex DNA. When super-helical DNA is relaxed by prior incubation with topoisomerase, it is a poor substrate for catenation. Extensive homology stimulates the catenation of circular single-stranded DNA and superhelical DNA, whereas little reaction occurs between these forms of the closely related DNAs of phages φX174 and G4, indicating that, in conjunction with topoisomerase I, RecA protein can discriminate perfect or nearly perfect homology from a high degree of relatedness. Circular single-stranded G4 DNA reacts with superhelical DNA of a chimeric phage, M13Goril, to form catenanes, at least half of which survive heating at 80°C following restriction cleavage in the M13 region, but few of which survive following restriction cleavage in the G4 region. Electron microscopic examination of catenated molecules cleaved in the M13 region reveals that in most cases the single-stranded G4 DNA is joined to the linear duplex M13(G4) DNA in the homologous G4 region. The junction frequently has the appearance of a D loop, with an extent equivalent to 100 or more bp. We conclude that a significant fraction of catenanes were hemicatenanes, in which the single-stranded circle was topologically linked, probably by multiple turns, to its complementary strand in the duplex DNA. These observations support the previous conclusion that RecA protein can pair a single strand with its complementary strand in duplex DNA in a side-by-side fashion without a free end in any of the three strands.

Item Type:Article
Source:Copyright of this article belongs to Elsevier Science.
ID Code:65836
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