This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Ball, H. L.
Right arrow Articles by Cortez, D.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Ball, H. L.
Right arrow Articles by Cortez, D.

 Previous Article  |  Next Article 

Molecular and Cellular Biology, May 2007, p. 3367-3377, Vol. 27, No. 9
0270-7306/07/$08.00+0     doi:10.1128/MCB.02238-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Function of a Conserved Checkpoint Recruitment Domain in ATRIP Proteins{triangledown}

Heather L. Ball,1 Mark R. Ehrhardt,2 Daniel A. Mordes,1 Gloria G. Glick,1 Walter J. Chazin,2 and David Cortez1*

Department of Biochemistry,1 Departments of Biochemistry and Chemistry and Center for Structural Biology, Vanderbilt University, Nashville, Tennessee 372322

Received 29 November 2006/ Returned for modification 29 December 2006/ Accepted 16 February 2007

The ATR (ATM and Rad3-related) kinase is essential to maintain genomic integrity. ATR is recruited to DNA lesions in part through its association with ATR-interacting protein (ATRIP), which in turn interacts with the single-stranded DNA binding protein RPA (replication protein A). In this study, a conserved checkpoint protein recruitment domain (CRD) in ATRIP orthologs was identified by biochemical mapping of the RPA binding site in combination with nuclear magnetic resonance, mutagenesis, and computational modeling. Mutations in the CRD of the Saccharomyces cerevisiae ATRIP ortholog Ddc2 disrupt the Ddc2-RPA interaction, prevent proper localization of Ddc2 to DNA breaks, sensitize yeast to DNA-damaging agents, and partially compromise checkpoint signaling. These data demonstrate that the CRD is critical for localization and optimal DNA damage responses. However, the stimulation of ATR kinase activity by binding of topoisomerase binding protein 1 (TopBP1) to ATRIP-ATR can occur independently of the interaction of ATRIP with RPA. Our results support the idea of a multistep model for ATR activation that requires separable localization and activation functions of ATRIP.

* Corresponding author. Mailing address: Department of Biochemistry, Vanderbilt University, 613 Light Hall, 23rd @ Pierce Ave., Nashville, TN 37232. Phone: (615) 322-8547. Fax: (615) 343-0704. E-mail: david.cortez{at}vanderbilt.edu

{triangledown} Published ahead of print on 5 March 2007.

Molecular and Cellular Biology, May 2007, p. 3367-3377, Vol. 27, No. 9
0270-7306/07/$08.00+0     doi:10.1128/MCB.02238-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Lovejoy, C. A., Xu, X., Bansbach, C. E., Glick, G. G., Zhao, R., Ye, F., Sirbu, B. M., Titus, L. C., Shyr, Y., Cortez, D. (2009). Functional genomic screens identify CINP as a genome maintenance protein. Proc. Natl. Acad. Sci. USA 106: 19304-19309 [Abstract] [Full Text]  
  • Bansbach, C. E., Betous, R., Lovejoy, C. A., Glick, G. G., Cortez, D. (2009). The annealing helicase SMARCAL1 maintains genome integrity at stalled replication forks. Genes Dev. 23: 2405-2414 [Abstract] [Full Text]  
  • Yan, S., Michael, W. M. (2009). TopBP1 and DNA polymerase-{alpha} directly recruit the 9-1-1 complex to stalled DNA replication forks. JCB 184: 793-804 [Abstract] [Full Text]  
  • Shiotani, B., Zou, L. (2009). ATR signaling at a glance. J. Cell Sci. 122: 301-304 [Full Text]  
  • Xu, X., Vaithiyalingam, S., Glick, G. G., Mordes, D. A., Chazin, W. J., Cortez, D. (2008). The Basic Cleft of RPA70N Binds Multiple Checkpoint Proteins, Including RAD9, To Regulate ATR Signaling. Mol. Cell. Biol. 28: 7345-7353 [Abstract] [Full Text]  
  • Binz, S. K., Wold, M. S. (2008). Regulatory Functions of the N-terminal Domain of the 70-kDa Subunit of Replication Protein A (RPA). J. Biol. Chem. 283: 21559-21570 [Abstract] [Full Text]  
  • Mordes, D. A., Glick, G. G., Zhao, R., Cortez, D. (2008). TopBP1 activates ATR through ATRIP and a PIKK regulatory domain. Genes Dev. 22: 1478-1489 [Abstract] [Full Text]  
  • Choi, J.-H., Lindsey-Boltz, L. A., Sancar, A. (2007). Reconstitution of a human ATR-mediated checkpoint response to damaged DNA. Proc. Natl. Acad. Sci. USA 104: 13301-13306 [Abstract] [Full Text]  
  • Delacroix, S., Wagner, J. M., Kobayashi, M., Yamamoto, K.-i., Karnitz, L. M. (2007). The Rad9-Hus1-Rad1 (9-1-1) clamp activates checkpoint signaling via TopBP1. Genes Dev. 21: 1472-1477 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»