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GENES & DEVELOPMENT 20:3104-3116, 2006
©2006 by Cold Spring Harbor Laboratory Press; ISSN 0890-9369/ $5.00
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The S. cerevisiae Rrm3p DNA helicase moves with the replication fork and affects replication of all yeast chromosomes

Anna Azvolinsky1, Stephen Dunaway1, Jorge Z. Torres, Jessica B. Bessler, and Virginia A. Zakian2

Department of Molecular Biology, Princeton University, Princeton, New Jersey 08544, USA

The Saccharomyces cerevisiae DNA helicase Rrm3p is needed for normal fork progression through >1000 discrete sites scattered throughout the genome. Here we show that replication of all yeast chromosomes was markedly delayed in rrm3 cells. Delayed replication was seen even in a region that lacks any predicted Rrm3p-dependent sites. Based on the pattern of replication intermediates in two-dimensional gels, the rate of fork movement in rrm3 cells appeared similar to wild-type except at known Rrm3p-dependent sites. These data suggest that although Rrm3p has a global role in DNA replication, its activity is needed only or primarily at specific, difficult-to-replicate sites. By the criterion of chromatin immunoprecipitation, Rrm3p was associated with both Rrm3p-dependent and -independent sites, and moved with the replication fork through both. In addition, Rrm3p interacted with Pol2p, the catalytic subunit of DNA polymerase {varepsilon}, in vivo. Thus, rather than being recruited to its sites of action when replication forks stall at these sites, Rrm3p is likely a component of the replication fork apparatus.

[Keywords: Rrm3p; Mrc1p; DNA replication; helicase; yeast; chromatin]

Received August 2, 2006; revised version accepted September 19, 2006.

1 These authors contributed equally to this work.

2 Corresponding author.

E-MAIL vzakian{at}princeton.edu; FAX (609) 258-1701.

Supplemental material is available at http://www.genesdev.org.

Article is online at http://www.genesdev.org/cgi/doi/10.1101/gad.1478906


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