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Molecular and Cellular Biology, June 2004, p. 5130-5143, Vol. 24, No. 12
0270-7306/04/$08.00+0     DOI: 10.1128/MCB.24.12.5130-5143.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

A New Saccharomyces cerevisiae Strain with a Mutant Smt3-Deconjugating Ulp1 Protein Is Affected in DNA Replication and Requires Srs2 and Homologous Recombination for Its Viability

Christine Soustelle ,1,{dagger},{ddagger} Laurence Vernis,2,{dagger} Karine Fréon,2 Anne Reynaud-Angelin,2 Roland Chanet,2 Francis Fabre,1 and Martine Heude2*

Commissariat à l'Energie Atomique, UMR 217 CNRS-CEA, DSV/DRR, 92265 Fontenay-aux-Roses,1 Institut Curie, UMR 2027/CNRS-Institut Curie, Génotoxicologie et Cycle Cellulaire, Centre Universitaire, 91405 Orsay Cedex, France2

Received 24 December 2003/ Returned for modification 25 February 2004/ Accepted 22 March 2004

The Saccharomyces cerevisiae Srs2 protein is involved in DNA repair and recombination. In order to gain better insight into the roles of Srs2, we performed a screen to identify mutations that are synthetically lethal with an srs2 deletion. One of them is a mutated allele of the ULP1 gene that encodes a protease specifically cleaving Smt3-protein conjugates. This allele, ulp1-I615N, is responsible for an accumulation of Smt3-conjugated proteins. The mutant is unable to grow at 37°C. At permissive temperatures, it still shows severe growth defects together with a strong hyperrecombination phenotype and is impaired in meiosis. Genetic interactions between ulp1 and mutations that affect different repair pathways indicated that the RAD51-dependent homologous recombination mechanism, but not excision resynthesis, translesion synthesis, or nonhomologous end-joining processes, is required for the viability of the mutant. Thus, both Srs2, believed to negatively control homologous recombination, and the process of recombination per se are essential for the viability of the ulp1 mutant. Upon replication, mutant cells accumulate single-stranded DNA interruptions. These structures are believed to generate different recombination intermediates. Some of them are fixed by recombination, and others require Srs2 to be reversed and fixed by an alternate pathway.

* Corresponding author. Mailing address: Institut Curie, UMR 2027/CNRS-Institut Curie, Génotoxicologie et Cycle Cellulaire, BÂtiment 110, Centre Universitaire, 91405 Orsay Cedex, France. Phone: (33) 1 69 86 30 56. Fax: (33) 1 69 86 94 29. E-mail: martine.heude{at}curie.u-psud.fr.

{dagger} C.S. and L.V. contributed equally to this work.

{ddagger} Present address: CNRS UPR 2167, Centre de Génétique Moléculaire, Equipe LIPM, 91198 Gif-sur-Yvette, France.

Molecular and Cellular Biology, June 2004, p. 5130-5143, Vol. 24, No. 12
0022-538X/04/$08.00+0     DOI: 10.1128/MCB.24.12.5130-5143.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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