Unwinding of the nascent lagging strand by Rep and PriA enables direct restart of stalled replication forks

doi:10.1074/jbc.M507224200

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Papers In Press, published online ahead of print August 2, 2005
J. Biol. Chem, 10.1074/jbc.M507224200
Submitted on July 5, 2005
Revised on August 2, 2005
Accepted on August 2, 2005

Unwinding of the nascent lagging strand by Rep and PriA enables direct restart of stalled replication forks

Ryan C. Heller and Kenneth J. Marians

Molecular Biology Program, Memorial Sloan-Kettering Cancer Center, New York, NY 10021

Corresponding Author: k-marians{at}ski.mskcc.org

During origin-independent replisome assembly, the replication restart protein PriC prefers to load the replication fork helicase, DnaB, to stalled replication forks where there is a gap in the nascent leading strand. However, this activity can be obstructed if the 5'-end of the nascent lagging strand is near the template branch point. Here we provide biochemical evidence that the helicase activities of Rep and PriA function to unwind the nascent lagging-strand DNA at such stalled replication forks. PriC then loads the replicative helicase, DnaB, onto the newly-generated, single-stranded template for the purposes of replisome assembly and duplex unwinding ahead of the replication fork. Direct rescue of replication forks by the Rep-PriC and PriA-PriC pathways in this manner may contribute to genomic stability by avoiding the potential dangers of fork breakage inherent to recombination-dependent restart pathways.

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