Nucleic Acids Research, 2000, Vol. 28, No. 19 3725-3732
© 2000 Oxford University Press
Phosphorylation of the replication protein A large subunit in the Saccharomyces cerevisiae checkpoint response
Department of Molecular Biology and Genetics, The Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA
The checkpoint mechanisms that delay cell cycle progression in response to DNA damage or inhibition of DNA replication are necessary for maintenance of genetic stability in eukaryotic cells. Potential targets of checkpoint-mediated regulation include proteins directly involved in DNA metabolism, such as the cellular single-stranded DNA (ssDNA) binding protein, replication protein A (RPA). Studies in Saccharomyces cerevisiae have revealed that the RPA large subunit (Rfa1p) is involved in the G1 and S phase DNA damage checkpoints. We now demonstrate that Rfa1p is phosphorylated in response to various forms of genotoxic stress, including radiation and hydroxyurea exposure, and further show that phosphorylation of Rfa1p is dependent on the central checkpoint regulator Mec1p. Analysis of the requirement for other checkpoint genes indicates that different mechanisms mediate radiation- and hydroxyurea-induced Rfa1p phosphorylation despite the common requirement for functional Mec1p. In addition, experiments with mutants defective in the Cdc13p telomere-binding protein indicate that ssDNA formation is an important signal for Rfa1p phosphorylation. Because Rfa1p contains the major ssDNA binding activity of the RPA heterotrimer and is required for DNA replication, repair and recombination, it is possible that phosphorylation of this subunit is directly involved in modulating RPA activity during the checkpoint response.
* To whom correspondence should be addressed at present address: Program in Molecular Biology and Genetics, Karmanos Cancer Institute, Wayne State University, Detroit, MI 48201, USA. Tel: +1 313 833 0715; Fax: +1 313 832 7294; Email: brushg@karmanos.org
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  S. Dulev, C. de Renty, R. Mehta, I. Minkov, E. Schwob, and A. Strunnikov Essential global role of CDC14 in DNA synthesis revealed by chromosome underreplication unrecognized by checkpoints in cdc14 mutants PNAS, August 25, 2009; 106(34): 14466 - 14471. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 V. M. Navadgi-Patil and P. M. Burgers Yeast DNA Replication Protein Dpb11 Activates the Mec1/ATR Checkpoint Kinase J. Biol. Chem., December 19, 2008; 283(51): 35853 - 35859. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
E. Olson, C. J. Nievera, V. Klimovich, E. Fanning, and X. Wu RPA2 Is a Direct Downstream Target for ATR to Regulate the S-phase Checkpoint J. Biol. Chem., December 22, 2006; 281(51): 39517 - 39533. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 K. Herzberg, V. I. Bashkirov, M. Rolfsmeier, E. Haghnazari, W. H. McDonald, S. Anderson, E. V. Bashkirova, J. R. Yates III, and W.-D. Heyer Phosphorylation of Rad55 on Serines 2, 8, and 14 Is Required for Efficient Homologous Recombination in the Recovery of Stalled Replication Forks Mol. Cell. Biol., November 15, 2006; 26(22): 8396 - 8409. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. J. Bartrand, D. Iyasu, S. M. Marinco, and G. S. Brush Evidence of Meiotic Crossover Control in Saccharomyces cerevisiae Through Mec1-Mediated Phosphorylation of Replication Protein A Genetics, January 1, 2006; 172(1): 27 - 39. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 T. M. Roberts, M. S. Kobor, S. A. Bastin-Shanower, M. Ii, S. A. Horte, J. W. Gin, A. Emili, J. Rine, S. J. Brill, and G. W. Brown Slx4 Regulates DNA Damage Checkpoint-dependent Phosphorylation of the BRCT Domain Protein Rtt107/Esc4 Mol. Biol. Cell, January 1, 2006; 17(1): 539 - 548. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. A. Cobb, T. Schleker, V. Rojas, L. Bjergbaek, J. A. Tercero, and S. M. Gasser Replisome instability, fork collapse, and gross chromosomal rearrangements arise synergistically from Mec1 kinase and RecQ helicase mutations Genes & Dev., December 15, 2005; 19(24): 3055 - 3069. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. Nakada, Y. Hirano, Y. Tanaka, and K. Sugimoto Role of the C Terminus of Mec1 Checkpoint Kinase in Its Localization to Sites of DNA Damage Mol. Biol. Cell, November 1, 2005; 16(11): 5227 - 5235. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
E. Bochkareva, L. Kaustov, A. Ayed, G.-S. Yi, Y. Lu, A. Pineda-Lucena, J. C. C. Liao, A. L. Okorokov, J. Milner, C. H. Arrowsmith, et al. Single-stranded DNA mimicry in the p53 transactivation domain interaction with replication protein A PNAS, October 25, 2005; 102(43): 15412 - 15417. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. J. Bartrand, D. Iyasu, and G. S. Brush DNA Stimulates Mec1-mediated Phosphorylation of Replication Protein A J. Biol. Chem., June 18, 2004; 279(25): 26762 - 26767. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. J. Millership, X. Cai, and G. Zhu Functional characterization of replication protein A2 (RPA2) from Cryptosporidium parvum Microbiology, May 1, 2004; 150(5): 1197 - 1205. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. G. Alexandrow and J. L. Hamlin Cdc6 Chromatin Affinity Is Unaffected by Serine-54 Phosphorylation, S-Phase Progression, and Overexpression of Cyclin A Mol. Cell. Biol., February 15, 2004; 24(4): 1614 - 1627. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S.-J. Park, S. L. M. Ciccone, B. Freie, A. Kurimasa, D. J. Chen, G. C. Li, D. W. Clapp, and S.-H. Lee A Positive Role for the Ku Complex in DNA Replication Following Strand Break Damage in Mammals J. Biol. Chem., February 13, 2004; 279(7): 6046 - 6055. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. M. Clifford, S. M. Marinco, and G. S. Brush The Meiosis-specific Protein Kinase Ime2 Directs Phosphorylation of Replication Protein A J. Biol. Chem., February 13, 2004; 279(7): 6163 - 6170. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Y. Ishimi, Y. Komamura-Kohno, H.-J. Kwon, K. Yamada, and M. Nakanishi Identification of MCM4 as a Target of the DNA Replication Block Checkpoint System J. Biol. Chem., June 27, 2003; 278(27): 24644 - 24650. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
N. Kantake, T. Sugiyama, R. D. Kolodner, and S. C. Kowalczykowski The Recombination-deficient Mutant RPA (rfa1-t11) Is Displaced Slowly from Single-stranded DNA by Rad51 Protein J. Biol. Chem., June 20, 2003; 278(26): 23410 - 23417. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 L. Zou and S. J. Elledge Sensing DNA Damage Through ATRIP Recognition of RPA-ssDNA Complexes Science, June 6, 2003; 300(5625): 1542 - 1548. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. M. Nakamura, B. A. Moser, and P. Russell Telomere Binding of Checkpoint Sensor and DNA Repair Proteins Contributes to Maintenance of Functional Fission Yeast Telomeres Genetics, August 1, 2002; 161(4): 1437 - 1452. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 F. Dantzer, L. Luna, M. Bjoras, and E. Seeberg Human OGG1 undergoes serine phosphorylation and associates with the nuclear matrix and mitotic chromatin in vivo Nucleic Acids Res., June 1, 2002; 30(11): 2349 - 2357. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
G. S. Brush, D. M. Clifford, S. M. Marinco, and A. J. Bartrand Replication protein A is sequentially phosphorylated during meiosis Nucleic Acids Res., December 1, 2001; 29(23): 4808 - 4817. [Abstract] [Full Text] [PDF]
|
|