Molecular and Cellular Biology, May 2004, p. 4151-4165, Vol. 24, No. 10
0270-7306/04/$08.00+0 DOI: 10.1128/MCB.24.10.4151-4165.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.
The Functions of Budding Yeast Sae2 in the DNA Damage Response Require Mec1- and Tel1-Dependent Phosphorylation
Enrico Baroni,
Valeria Viscardi,
Hugo Cartagena-Lirola, Giovanna Lucchini, and Maria Pia Longhese*
Dipartimento di Biotecnologie e Bioscienze, Università degli Studi di Milano-Bicocca, 20126 Milan, Italy
Received 30 October 2003/
Returned for modification 23 December 2003/
Accepted 9 February 2004
DNA damage checkpoint pathways sense DNA lesions and transduce the signals into appropriate biological responses, including cell cycle arrest, induction of transcriptional programs, and modification or activation of repair factors. Here we show that the Saccharomyces cerevisiae Sae2 protein, known to be involved in processing meiotic and mitotic double-strand breaks, is required for proper recovery from checkpoint-mediated cell cycle arrest after DNA damage and is phosphorylated periodically during the unperturbed cell cycle and in response to DNA damage. Both cell cycle- and DNA damage-dependent Sae2 phosphorylation requires the main checkpoint kinase, Mec1, and the upstream components of its pathway, Ddc1, Rad17, Rad24, and Mec3. Another pathway, involving Tel1 and the MRX complex, is also required for full DNA damage-induced Sae2 phosphorylation, that is instead independent of the downstream checkpoint transducers Rad53 and Chk1, as well as of their mediators Rad9 and Mrc1. Mutations altering all the favored ATM/ATR phosphorylation sites of Sae2 not only abolish its in vivo phosphorylation after DNA damage but also cause hypersensitivity to methyl methanesulfonate treatment, synthetic lethality with RAD27 deletion, and decreased rates of mitotic recombination between inverted Alu repeats, suggesting that checkpoint-mediated phosphorylation of Sae2 is important to support its repair and recombination functions.
* Corresponding author. Mailing address: Dipartimento di Biotecnologie e Bioscienze, Università degli Studi di Milano-Bicocca, P.zza della Scienza 2, 20126 Milan, Italy. Phone: 39-02-64483425. Fax: 39-02-64483565. E-mail: mariapia.longhese{at}unimib.it.
E.B. and V.V. contributed equally to this work.
Molecular and Cellular Biology, May 2004, p. 4151-4165, Vol. 24, No. 10
0022-538X/04/$08.00+0 DOI: 10.1128/MCB.24.10.4151-4165.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.
This article has been cited by other articles:
-
Akamatsu, Y., Murayama, Y., Yamada, T., Nakazaki, T., Tsutsui, Y., Ohta, K., Iwasaki, H.
(2008). Molecular Characterization of the Role of the Schizosaccharomyces pombe nip1+/ctp1+ Gene in DNA Double-Strand Break Repair in Association with the Mre11-Rad50-Nbs1 Complex. Mol. Cell. Biol.
28: 3639-3651
[Abstract]
[Full Text]
-
Kim, H.-S., Vijayakumar, S., Reger, M., Harrison, J. C., Haber, J. E., Weil, C., Petrini, J. H. J.
(2008). Functional Interactions Between Sae2 and the Mre11 Complex. Genetics
178: 711-723
[Abstract]
[Full Text]
-
Lee, K., Lee, S. E.
(2007). Saccharomyces cerevisiae Sae2- and Tel1-Dependent Single-Strand DNA Formation at DNA Break Promotes Microhomology-Mediated End Joining. Genetics
176: 2003-2014
[Abstract]
[Full Text]
-
Kitagawa, T., Hoshida, H., Akada, R.
(2007). Genome-Wide Analysis of Cellular Response to Bacterial Genotoxin CdtB in Yeast. Infect. Immun.
75: 1393-1402
[Abstract]
[Full Text]
-
Clerici, M., Mantiero, D., Lucchini, G., Longhese, M. P.
(2005). The Saccharomyces cerevisiae Sae2 Protein Promotes Resection and Bridging of Double Strand Break Ends. J. Biol. Chem.
280: 38631-38638
[Abstract]
[Full Text]
-
Rattray, A. J., Shafer, B. K., Neelam, B., Strathern, J. N.
(2005). A mechanism of palindromic gene amplification in Saccharomyces cerevisiae. Genes Dev.
19: 1390-1399
[Abstract]
[Full Text]
-
Clerici, M., Baldo, V., Mantiero, D., Lottersberger, F., Lucchini, G., Longhese, M. P.
(2004). A Tel1/MRX-Dependent Checkpoint Inhibits the Metaphase-to-Anaphase Transition after UV Irradiation in the Absence of Mec1. Mol. Cell. Biol.
24: 10126-10144
[Abstract]
[Full Text]
Copyright © 2004 by the American Society for Microbiology. All rights reserved.