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

 Previous Article  |  Next Article 

Molecular and Cellular Biology, April 2004, p. 3277-3285, Vol. 24, No. 8
0270-7306/04/$08.00+0     DOI: 10.1128/MCB.24.8.3277-3285.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Association of Rad9 with Double-Strand Breaks through a Mec1-Dependent Mechanism

Division of Biological Science, Graduate School of Science, Nagoya University, Chikusa-ku, Nagoya 464-0814, Japan

Received 10 September 2003/ Returned for modification 4 November 2003/ Accepted 19 January 2004

Rad9 is required for the activation of DNA damage checkpoint pathways in budding yeast. Rad9 is phosphorylated after DNA damage in a Mec1- and Tel1-dependent manner and subsequently interacts with Rad53. This Rad9-Rad53 interaction has been suggested to trigger the activation and phosphorylation of Rad53. Here we show that Mec1 controls the Rad9 accumulation at double-strand breaks (DSBs). Rad9 was phosphorylated after DSB induction and associated with DSBs. However, its phosphorylation and association with DSBs were significantly decreased in cells carrying a mec1 or kinase-negative mec1 mutation. Mec1 phosphorylated the S/TQ motifs of Rad9 in vitro, the same motifs that are phosphorylated after DNA damage in vivo. In addition, multiple mutations in the Rad9 S/TQ motifs resulted in its defective association with DSBs. Phosphorylation of Rad9 was partially defective in cells carrying a weak mec1 allele (mec1-81), whereas its association with DSBs occurred efficiently in the mec1-81 mutants, as found in wild-type cells. However, the Rad9-Rad53 interaction after DSB induction was significantly decreased in mec1-81 mutants, as it was in mec1 mutants. Deletion mutation in RAD53 did not affect the association of Rad9 with DSBs. Our results suggest that Mec1 promotes association of Rad9 with sites of DNA damage, thereby leading to full phosphorylation of Rad9 and its interaction with Rad53.

This article has been cited by other articles:

• Grandin, N., Charbonneau, M. (2007). Control of the yeast telomeric senescence survival pathways of recombination by the Mec1 and Mec3 DNA damage sensors and RPA. Nucleic Acids Res 35: 822-838 [Abstract] [Full Text]  
• Tseng, S.-F., Lin, J.-J., Teng, S.-C. (2006). The telomerase-recruitment domain of the telomere binding protein Cdc13 is regulated by Mec1p/Tel1p-dependent phosphorylation. Nucleic Acids Res 34: 6327-6336 [Abstract] [Full Text]  
• Ma, J.-L., Lee, S.-J., Duong, J. K., Stern, D. F. (2006). Activation of the Checkpoint Kinase Rad53 by the Phosphatidyl Inositol Kinase-like Kinase Mec1. J. Biol. Chem. 281: 3954-3963 [Abstract] [Full Text]  
• Nakada, D., Hirano, Y., Tanaka, Y., Sugimoto, K. (2005). Role of the C Terminus of Mec1 Checkpoint Kinase in Its Localization to Sites of DNA Damage. Mol. Biol. Cell 16: 5227-5235 [Abstract] [Full Text]  
• Wysocki, R., Javaheri, A., Allard, S., Sha, F., Cote, J., Kron, S. J. (2005). Role of Dot1-Dependent Histone H3 Methylation in G1 and S Phase DNA Damage Checkpoint Functions of Rad9. Mol. Cell. Biol. 25: 8430-8443 [Abstract] [Full Text]  
• Giannattasio, M., Lazzaro, F., Plevani, P., Muzi-Falconi, M. (2005). The DNA Damage Checkpoint Response Requires Histone H2B Ubiquitination by Rad6-Bre1 and H3 Methylation by Dot1. J. Biol. Chem. 280: 9879-9886 [Abstract] [Full Text]  
• Lee, S.-J., Duong, J. K., Stern, D. F. (2004). A Ddc2-Rad53 Fusion Protein Can Bypass the Requirements for RAD9 and MRC1 in Rad53 Activation. Mol. Biol. Cell 15: 5443-5455 [Abstract] [Full Text]  
• Nakada, D., Hirano, Y., Sugimoto, K. (2004). Requirement of the Mre11 Complex and Exonuclease 1 for Activation of the Mec1 Signaling Pathway. Mol. Cell. Biol. 24: 10016-10025 [Abstract] [Full Text]