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Molecular and Cellular Biology, December 2005, p. 10721-10730, Vol. 25, No. 24
0270-7306/05/$08.00+0     doi:10.1128/MCB.25.24.10721-10730.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Cooperative Control of Crb2 by ATM Family and Cdc2 Kinases Is Essential for the DNA Damage Checkpoint in Fission Yeast

Toru M. Nakamura,^1,3* Bettina A. Moser,^1,3 Li-Lin Du,¹ and Paul Russell^1,2*

Departments of Molecular Biology,¹ Cell Biology, The Scripps Research Institute, La Jolla, California 92037,² Department of Biochemistry and Molecular Genetics, University of Illinois at Chicago, Chicago, Illinois 60607³

Received 29 August 2005/ Returned for modification 19 September 2005/ Accepted 26 September 2005

The cellular responses to double-stranded breaks (DSBs) typically involve the extensive accumulation of checkpoint proteins in chromatin surrounding the damaged DNA. One well-characterized example involves the checkpoint protein Crb2 in the fission yeast Schizosaccharomyces pombe. The accumulation of Crb2 at DSBs requires the C-terminal phosphorylation of histone H2A (known as -H2A) by ATM family kinases in chromatin surrounding the break. It also requires the constitutive methylation of histone H4 on lysine-20 (K20). Interestingly, neither type of histone modification is essential for the Crb2-dependent checkpoint response. However, H4-K20 methylation is essential in a crb2-T215A strain that lacks a cyclin-dependent kinase phosphorylation site in Crb2. Here we explain this genetic interaction by describing a previously overlooked effect of the crb2-T215A mutation. We show that crb2-T215A cells are able to initiate but not sustain a checkpoint response. We also report that -H2A is essential for the DNA damage checkpoint in crb2-T215A cells. Importantly, we show that inactivation of Cdc2 in -H2A-defective cells impairs Crb2-dependent signaling to the checkpoint kinase Chk1. These findings demonstrate that full Crb2 activity requires phosphorylation of threonine-215 by Cdc2. This regulation of Crb2 is independent of the histone modifications that are required for the hyperaccumulation of Crb2 at DSBs.

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* Corresponding author. Mailing address for Paul Russell: Department of Molecular Biology, MB3, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, CA 92037. Phone: (858) 784-8273. Fax: (858) 784-2265. E-mail: prussell{at}scripps.edu. Mailing address for Toru M. Nakamura: Department of Biochemistry and Molecular Genetics, University of Illinois at Chicago, 900 S. Ashland Ave. M/C669, Chicago, IL 60607. Phone: (312) 996-1988. Fax: (312) 413-0353. E-mail: nakamut{at}uic.edu.

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Molecular and Cellular Biology, December 2005, p. 10721-10730, Vol. 25, No. 24
0022-538X/05/$08.00+0     doi:10.1128/MCB.25.24.10721-10730.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

This article has been cited by other articles:

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