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Papers In Press, published online ahead of print September 10, 2001
Institute of Genetics, University of Nottingham, Nottingham NG7 2UH
Corresponding Author: bob.lloyd{at}nottingham.ac.uk
The replicative apparatus often encounters blocks to its progression that necessitate removal of the block and reloading of the replication machinery. In Escherichia coli a major pathway of replication restart involves unwinding of the stalled fork to generate a four-stranded Holliday junction which can then be cleaved by the RuvABC helicase/endonuclease. This fork regression may be catalysed by RecG but is thought to occur even in its absence. Here we test whether RuvAB helicase can also catalyse unwinding of forked DNA to form Holliday junctions. We find that fork DNA is unwound in the direction required for Holliday junction formation only if loading of RuvB is restricted to the parental duplex DNA arm. If binding of RuvB is unrestricted then RuvAB preferentially unwinds forks in the opposing direction. This is likely related to the greater efficiency of two opposed RuvB hexamers operating across a junction compared with a single hexamer. These data argue against RuvAB acting directly at damaged replication forks and imply that other mechanisms must operate in vivo to catalyse Holliday junction formation.
J. Biol. Chem, 10.1074/jbc.M107945200
Submitted on August 17, 2001
Revised on September 10, 2001
Accepted on September 9, 2001
Action of RuvAB at replication fork structures
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