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Characterization of mutations that are synthetic lethal with pol3-13, a mutated allele of DNA polymerase delta in Saccharomyces cerevisiae

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Abstract

The pol3-13 mutation is located in the C-terminal end of POL3, the gene encoding the catalytic subunit of polymerase δ, and confers thermosensitivity onto the Saccharomyces cerevisiae mutant strain. To get insight about DNA replication control, we performed a genetic screen to identify genes that are synthetic lethal with pol3-13. Mutations in genes encoding the two other subunits of DNA polymerase δ (HYS2, POL32) were identified. Mutations in two recombination genes (RAD50, RAD51) were also identified, confirming that homologous recombination is necessary for pol3-13 mutant strain survival. Other mutations were identified in genes involved in repair and genome stability (MET18/MMS19), in the control of origin-firing and/or transcription (ABF1, SRB7), in the S/G2 checkpoint (RAD53), in the Ras-cAMP signal transduction pathway (MKS1), in nuclear pore metabolism (SEH1), in protein degradation (DOC1) and in folding (YDJ1). Finally, mutations in three genes of unknown function were isolated (NBP35, DRE2, TAH18). Synthetic lethality between pol3-13 and each of the three mutants pol32, mms19 and doc1 could be suppressed by a rad18 deletion, suggesting an important role of ubiquitination in DNA replication control. We propose that the pol3-13 mutant generates replicative problems that need both homologous recombination and an intact checkpoint machinery to be overcome.

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Acknowledgements

Continual interest by G. Faye is gratefully acknowledged. We are grateful to Z. Hrisoho, G. Cheret, B. Agoutin and K. Freon for excellent technical assistance. G. Baldacci, L. Vernis-Beringue and all the members of the UMR2027 group are acknowledged for their useful criticisms of this manuscript and for continuous discussions. We also thank Dr. L. Sperling for a thorough review of this manuscript. This work was supported by the Association pour la Recherche sur le Cancer (contracts 9747, 5862).

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  1. Institut Curie Recherche, CNRS UMR2027/Institut Curie, Bât. 110, Centre Universitaire, 91405, Orsay Cédex, France

    Roland Chanet & Martine Heude

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Chanet, R., Heude, M. Characterization of mutations that are synthetic lethal with pol3-13, a mutated allele of DNA polymerase delta in Saccharomyces cerevisiae . Curr Genet 43, 337–350 (2003). https://doi.org/10.1007/s00294-003-0407-2

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