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RESEARCH PAPER
1 Lawrence Berkeley National Laboratory and Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, California 94720, USA; 2 Department of Biochemistry and Molecular Biophysics, Washington University School of Medicine, St. Louis, Missouri 63110, USA
Chromatin assembly and DNA replication are temporally coupled, and DNA replication in the absence of histone synthesis causes inviability. Here we demonstrate that chromatin assembly factor Asf1 also affects DNA replication. In budding yeast cells lacking Asf1, the amounts of several DNA replication proteins, including replication factor C (RFC), proliferating cell nuclear antigen (PCNA), and DNA polymerase 
(Pol ), are reduced at stalled replication forks. In contrast, DNA polymerase 
(Pol ) accumulates to higher than normal levels at stalled forks in asf1
cells. Using purified, recombinant proteins, we demonstrate that RFC directly binds Asf1 and can recruit Asf1 to DNA molecules in vitro. We conclude that histone chaperone protein Asf1 maintains a subset of replication elongation factors at stalled replication forks and directly interacts with the replication machinery.
[Keywords: Chromatin assembly; histone deposition; DNA replication; genome stability]
Received February 11, 2005; revised version accepted April 12, 2005.
Article published online ahead of print. Article and publication date are at http://www.genesdev.org/cgi/doi/10.1101/gad.1305005.
3 Present address: Department of Biology, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
4 Address as of July 1, 2005: Program in Gene Function and Expression, University of Massachusetts Medical School, Worcester, MA 01605, USA.
5 Corresponding author.
E-MAIL pdkaufman{at}lbl.gov; FAX (510) 486-6488.
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