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Rad17 Plays a Central Role in Establishment of the Interaction between TopBP1 and the Rad9-Hus1-Rad1 Complex at Stalled Replication Forks

Lee, Joon and Dunphy, William G. (2010) Rad17 Plays a Central Role in Establishment of the Interaction between TopBP1 and the Rad9-Hus1-Rad1 Complex at Stalled Replication Forks. Molecular Biology of the Cell, 21 (6). pp. 926-935. ISSN 1059-1524. PMCID PMC2836973.

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Rad17 is critical for the ATR-dependent activation of Chk1 during checkpoint responses. It is known that Rad17 loads the Rad9-Hus1-Rad1 (9-1-1) complex onto DNA. We show that Rad17 also mediates the interaction of 9-1-1 with the ATR-activating protein TopBP1 in Xenopus egg extracts. Studies with Rad17 mutants indicate that binding of ATP to Rad17 is essential for the association of 9-1-1 and TopBP1. Furthermore, hydrolysis of ATP by Rad17 is necessary for the loading of 9-1-1 onto DNA and the elevated, checkpoint-dependent accumulation of TopBP1 on chromatin. Significantly, a mutant 9-1-1 complex that cannot bind TopBP1 has a normal capacity to promote elevated accumulation of TopBP1 on chromatin. Taken together, we propose the following mechanism. First, Rad17 loads 9-1-1 onto DNA. Second, TopBP1 accumulates on chromatin in a manner that depends on both Rad17 and 9-1-1. Finally, 9-1-1 and TopBP1 dock in a Rad17-dependent manner before activation of Chk1.

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Dunphy, William G.0000-0001-7598-8939
Additional Information:© 2010 by The American Society for Cell Biology. Under the License and Publishing Agreement, authors grant to the general public, effective two months after publication of (i.e.,. the appearance of) the edited manuscript in an online issue of MBoC, the nonexclusive right to copy, distribute, or display the manuscript subject to the terms of the Creative Commons–Noncommercial–Share Alike 3.0 Unported license ( Submitted November 17, 2009; Revised January 12, 2010; Accepted January 19, 2010. Monitoring Editor: Daniel J. Lew. This was published online ahead of print in MBC in Press ( on January 28, 2010. We are grateful to Drs. Karlene Cimprich (Stanford University), Jerard Hurwitz (Memorial Sloan-Kettering Cancer Center), and Howard Lindsay (Lancaster University, Lancaster, United Kingdom) for kindly supplying key reagents. We also thank other members of the laboratory for fruitful comments on this manuscript. This work was supported by grants from the National Institutes of Health (GM043974 and GM070891) to W.G.D.
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Issue or Number:6
PubMed Central ID:PMC2836973
Record Number:CaltechAUTHORS:20100406-101055081
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Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:17865
Deposited By: Tony Diaz
Deposited On:21 Apr 2010 19:13
Last Modified:09 Mar 2020 13:18

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