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GENES & DEVELOPMENT 21:898-903, 2007
©2007 by Cold Spring Harbor Laboratory Press; ISSN 0890-9369/ $5.00
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RESEARCH COMMUNICATION

The structural determinants of checkpoint activation

Christina A. MacDougall, Tony S. Byun, Christopher Van, Muh-ching Yee, and Karlene A. Cimprich1

Department of Chemical and Systems Biology, Stanford University, Stanford, California 94305, USA

Here, we demonstrate that primed, single-stranded DNA (ssDNA) is sufficient for activation of the ATR-dependent checkpoint pathway in Xenopus egg extracts. Using this structure, we define the contribution of the 5'- and 3'-primer ends to Chk1 activation when replication is blocked and ongoing. In addition, we show that although ssDNA is not sufficient for checkpoint activation, the amount of ssDNA adjacent to the primer influences the level of Chk1 phosphorylation. These observations define the minimal DNA requirements for checkpoint activation and suggest that primed ssDNA represents a common checkpoint activating-structure formed following many types of damage.

Received December 12, 2006; revised version accepted March 5, 2007.

1 Corresponding author.

E-MAIL cimprich{at}stanford.edu; FAX (650) 725-4665.

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

Article is online at http://www.genesdev.org/cgi/doi/10.1101/gad.1522607


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