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RecBCD Enzyme Switches Lead Motor Subunits in Response to χ Recognition
Journal article   Open access   Peer reviewed

RecBCD Enzyme Switches Lead Motor Subunits in Response to χ Recognition

Maria Spies, Ichiro Amitani, Ronald J. Baskin and Stephen C. Kowalczykowski
Cell, Vol.131(4), pp.694-705
2007
DOI: 10.1016/j.cell.2007.09.023
PMCID: PMC2151923
PMID: 18022364
url
https://doi.org/10.1016/j.cell.2007.09.023View
Published (Version of record) Open Access

Abstract

RecBCD is a DNA helicase comprising two motor subunits, RecB and RecD. Recognition of the recombination hotspot, χ, causes RecBCD to pause and reduce translocation speed. To understand this control of translocation, we used single-molecule visualization to compare RecBCD to the RecBCD K177Q mutant with a defective RecD motor. RecBCD K177Q paused at χ but did not change its translocation velocity. RecBCD K177Q translocated at the same rate as the wild-type post-χ enzyme, implicating RecB as the lead motor after χ. P1 nuclease treatment eliminated the wild-type enzyme's velocity changes, revealing a χ-containing ssDNA loop preceding χ recognition and showing that RecD is the faster motor before χ. We conclude that before χ, RecD is the lead motor but after χ, the slower RecB motor leads, implying a switch in motors at χ. We suggest that degradation of foreign DNA needs fast translocation, whereas DNA repair uses slower translocation to coordinate RecA loading onto ssDNA.
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