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The function of the secondary DNA‐binding site of RecA protein during DNA strand exchange
Journal article   Open access   Peer reviewed

The function of the secondary DNA‐binding site of RecA protein during DNA strand exchange

Alexander V Mazin and Stephen C Kowalczykowski
The EMBO journal, v 17(4), pp 1161-1168
15 Feb 1998
PMID: 9463393
url
https://doi.org/10.1093/emboj/17.4.1161View
Published, Version of Record (VoR) Open

Abstract

homologous pairing DNA–protein interactions genetic recombination
RecA protein features two distinct DNA‐binding sites. During DNA strand exchange, the primary site binds to single‐stranded DNA (ssDNA), forming the helical RecA nucleoprotein filament. The weaker secondary site binds double‐stranded DNA (dsDNA) during the homology search process. Here we demonstrate that this site has a second important function. It binds the ssDNA strand that is displaced from homologous duplex DNA during DNA strand exchange, stabilizing the initial heteroduplex DNA product. Although the high affinity of the secondary site for ssDNA is essential for DNA strand exchange, it renders DNA strand exchange sensitive to an excess of ssDNA which competes with dsDNA for binding. We further demonstrate that single‐stranded DNA‐binding protein can sequester ssDNA, preventing its binding to the secondary site and thereby assisting at two levels: it averts the inhibition caused by an excess of ssDNA and prevents the reversal of DNA strand exchange by removing the displaced strand from the secondary site.

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Web of Science research areas
Biochemistry & Molecular Biology
Cell Biology
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