J. Biol. Chem., Vol. 266, Issue 16, 10112-10121, 06, 1991

DNA substrate requirements for stable joint molecule formation by the RecA and single-stranded DNA-binding proteins of Escherichia coli [published erratum appears in J Biol Chem 1991 Oct 15;266(29):19865]

BB Konforti and RW Davis
Department of Biochemistry, Stanford University, California 94305-5307.

In reactions between linear single-stranded DNAs (ssDNAs) and circular double-stranded DNAs (dsDNAs), stable joint molecule formation promoted by the recA protein (RecA) requires negative superhelicity, a homologous end, and an RecA-ssDNA complex. Linear ssDNAs with 3'-end homology react more efficiently than linear ssDNAs with 5'-end homology. This 3'-end preference is explained by the finding that 3'- ends are more effectively coated by RecA than 5'-ends, as judged by exonuclease VII protection, and are thus more reactive. The ability of linear ssDNAs with 5'-end homology to react is improved by the presence of low concentrations of exonuclease VII. In reactions between ssDNAs and linear dsDNAs with end homology, stable joint molecule formation occurs more efficiently when the homology is at the 3'-end rather than at the 5'-end of the complementary strand. In addition, linear dsDNAs with homology at the 3'-end of the complementary strand react more efficiently with linear ssDNAs with 3'-end homology than with linear ssDNAs with 5'-end homology. The ability of linear ssDNAs with 5'-end homology to react, in the absence of single-stranded DNA-binding protein, is improved by adding 33-46 nucleotides of heterologous sequence to the 5'-end of the linear ssDNA. The poor reactivity of linear ssDNAs with 5'-end homology is explained by a lack of RecA at the 5'-ends of linear ssDNAs, which is a consequence of the polar association and dissociation of RecA.
CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				J. M. Bork, M. M. Cox, and R. B. Inman RecA Protein Filaments Disassemble in the 5' to 3' Direction on Single-stranded DNA J. Biol. Chem., November 30, 2001; 276(49): 45740 - 45743. [Abstract] [Full Text] [PDF]

				S. J. Shi, A. Scheffer, E. Bjeldanes, M. A. Reynolds, and L. J. Arnold DNA exhibits multi-stranded binding recognition on glass microarrays Nucleic Acids Res., October 15, 2001; 29(20): 4251 - 4256. [Abstract] [Full Text] [PDF]