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Molecular and Cellular Biology, November 2003, p. 7861-7874, Vol. 23, No. 21
0270-7306/03/$08.00+0     DOI: 10.1128/MCB.23.21.7861-7874.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Swi1 Prevents Replication Fork Collapse and Controls Checkpoint Kinase Cds1

Eishi Noguchi, Chiaki Noguchi, Li-Lin Du, and Paul Russell*

Department of Molecular Biology and Cell Biology, The Scripps Research Institute, La Jolla, California 92037

Received 8 May 2003/ Returned for modification 5 June 2003/ Accepted 30 July 2003

The replication checkpoint is a dedicated sensor-response system activated by impeded replication forks. It stabilizes stalled forks and arrests division, thereby preserving genome integrity and promoting cell survival. In budding yeast, Tof1 is thought to act as a specific mediator of the replication checkpoint signal that activates the effector kinase Rad53. Here we report studies of fission yeast Swi1, a Tof1-related protein required for a programmed fork-pausing event necessary for mating type switching. Our studies have shown that Swi1 is vital for proficient activation of the Rad53-like checkpoint kinase Cds1. Together they are required to prevent fork collapse in the ribosomal DNA repeats, and they also prevent irreversible fork arrest at a newly identified hydroxyurea pause site. Swi1 also has Cds1-independent functions. Rad22 DNA repair foci form during S phase in swi1 mutants and to a lesser extent in cds1 mutants, indicative of fork collapse. Mus81, a DNA endonuclease required for recovery from collapsed forks, is vital in swi1 but not cds1 mutants. Swi1 is recruited to chromatin during S phase. We propose that Swi1 stabilizes replication forks in a configuration that is recognized by replication checkpoint sensors.

* Corresponding author. Mailing address: Department of Molecular Biology and Cell Biology, MB-3, The Scripps Research Institute, 10550 North Torrey Pines Rd., La Jolla, CA 92037. Phone: (858) 784-8273. Fax: (858) 784-2265. E-mail: prussell{at}scripps.edu.

Molecular and Cellular Biology, November 2003, p. 7861-7874, Vol. 23, No. 21
0022-538X/03/$08.00+0     DOI: 10.1128/MCB.23.21.7861-7874.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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