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> Stages de M2 > stages proposés pour l’année 2019-2020 > Cibles de la polo-like kinase lors de la recombinaison méiotique

Cibles de la polo-like kinase lors de la recombinaison méiotique

proposé par Valérie Borde, Institut Curie – UMR3244 Dynamique de l’Information Génétique

Projet de stage :

Recombination can be due to accidental lesions, following exposure to genotoxic agents, or during replication failures, but lesions can also be deliberately self-inflicted by the cell, such as during meiosis, which is the process we study in the lab. In meiosis, programmed DNA double-strand breaks are formed by the Spo11 protein, and their repair by homologous recombination creates the crossovers, necessary for the correct segregation of homologs during the first meiotic division and for fertility. Crossover formation is highly controlled since it could potentially lead to harmful chromosomal rearrangements. In particular, crossover formation depends on the successful execution of other cell-cycle related events, and occurs only upon the induction of the polo-like kinase Cdc5. Cdc5 induction in meiosis is sufficient to promote crossover formation, but its targets in this process are unknown. The project will consist on one hand to perform a proteomic screen of Cdc5-associated proteins during meiosis, using conditions that are already optimized in the lab. After mass spectrometry analysis by the Curie Institute platform, the candidates will be confirmed by reverse pull-down, their phosphorylation status will be analyzed and phosphomutants will be designed to assess the consequence on crossover formation. In parallel, the student will also generate mutants for phosphosites that we have already identified in a candidate approach in one of the proteins involved in meiotic crossover formation.

Techniques mises en œuvre par le stagiaire :

The project will employ genetic, molecular biology and in vivo protein analyses, using the budding yeast as a model system.

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