Engineer Technician (M/F)

Laboratory

The Chromosome Dynamics and Recombination team is located at Institut Curie, one of the leading cancer research institutions. The institute provides access to high-quality technological platforms such as the High-throughput Sequencing and Proteomics platforms that will be useful to the project. Aurore Sanchez, CNRS researcher in the “Chromosome Dynamics and Recombination” team led by Valérie Borde, is looking for an engineer technician (M/F) in biology. The team is composed by one PI, a CNRS researcher, an engineer, 2 assistant engineers, a postdoctoral researcher and 2 PhD students (8 people in total). https://institut-curie.org/team/borde

Research subject of the team

The team studies homologous recombination and its importance for genome stability and chromosome segregation during the formation of haploid gametes in meiosis. We use genetic, genomic, proteomic and molecular biology approaches for our studies, and apply them to the yeast Saccharomyces cerevisiae as a model organism.

Recombination occurs when a chromosome that has undergone a double-stranded DNA break (DSB) uses a homologous sequence for its repair, resulting in the exchange of DNA between parental chromosomes at the origin of genetic diversity, called crossing-over. Recombination defects have severe clinical consequences, and are linked to several diseases, such as certain hereditary cancers, genetic instability syndromes and infertility in mammals. Although meiotic recombination has been studied for decades, many of the underlying molecular processes still unclear, largely because (i) many DSB repair players remain unknown, and (2) the known actors act at different times during homologous recombination making their identification or characterization a challenging task.

However, our recent development of an experimental system that allow to separate the different stages of homologous recombination couple with cutting edge technologies already set up or in development in the lab, might finally make te identification and characterization of novels and specific actors of each step possible.

Missions

The candidate work will consist in the identification of the different actors and their involvement in the formation of crossing-overs, using an experimental system that allows the temporal separation of the first steps of recombination from the later steps. The candidate's work will be to identify the actors of these recombination steps by proteomic screens, and to test their presence at recombination sites by chromatin immunoprecipitation but also their importance by analysing the phenotype of their inactivation in vivo (gametes viability, recombination efficiency, distribution and nature of recombination events, etc.). The candidate will use techniques routinely used in the team and will implement a global proteomic approach (technique derived from BioID) to identify new actors in the last steps of recombination.

Bibliographie :

1.        Ceppi, I., Dello Stritto, S., Mütze, MR., Braunshier, M., Mengoli, V., Reginato, G., Phúc Võ, HM., Jimeno, S., Acharya, A., Roy, M., Sanchez, A., Halder, S., Howard, SM., Guérois, R., Huertas, P., Noordermeer, MS., Seidel, R., and Cejka, P., 2024. Mechanism of BRCA1-BARD1 function in DNA end resection and DNA protection. Nature 634, pages 492–500 https://doi.org/10.1038/s41586-024-07909-9

2.        Legrand, S., Saifudeen, A., Bordelet, H., Vernerey, J., Guille A., Bignaud, A., Thierry, A., Acquaviva, L., Gaudin, M., Sanchez, A., Johnson, D., Friedrich, A., Schacherer, J., Neale, MJ., Borde, V.,Koszul, R., Llorente, B., 2024. Absence of chromosome axis protein recruitment prevents meiotic recombination chromosome-wide in the budding yeast Lachancea kluyveri. Proc. Natl. Acad. Sci. U. S. A. 121, e2312820121 https://doi.org/10.1073/pnas.2312820121

3.        Mengoli, V., Ceppi, I., Sanchez, A., Cannavo, E., Halder, S., Scaglione, S., Gaillard, PH., McHugh, P., Riesen, N., Pettazzoni, P., Cejka, P., 2022. WRN helicase and mismatch repair complexes independently and synergistically disrupt cruciform DNA structures. The EMBO Journal https://doi.org/10.15252/embj.2022111998

4.        Halder, S., Sanchez, A., Ranjha, L., Reginato, G., Ceppi, I., Acharya, A., Anand, R., Cejka, P., 2022. Double-stranded DNA binding function of RAD51 in DNA protection and its regulation by BRCA2. Mol Cell https://doi.org/10.1016/j.molcel.2022.08.014

5.        Dai*, J., Sanchez*, A., Adam, C., Ranjha, L., Reginato, G., Chervy, P., Tellier-Lebegue, C., Andreani, J., Guérois, R., Ropars, V., Du, M.-H.L., Maloisel, L., Martini, E., Legrand, P., Thureau, A., Cejka, P., Borde, V., Charbonnier, J.-B., 2021. Molecular basis of the dual role of the Mlh1-Mlh3 endonuclease in MMR and in meiotic crossover formation. Proc. Natl. Acad. Sci. 118. https://doi.org/10.1073/pnas.2022704118 *Equal contribution

6.        Cannavo*, E., Sanchez*, A., Anand*, R., Ranjha, L., Hugener, J., Adam, C., Acharya, A., Weyland, N., Aran-Guiu, X., Charbonnier, J.-B., Hoffmann, E.R., Borde, V., Matos, J., Cejka, P., 2020. Regulation of the MLH1–MLH3 endonuclease in meiosis. Nature 586, 618–622. https://doi.org/10.1038/s41586-020-2592-2 *Equal contribution

7.        Bennett, L.G., Wilkie, A.M., Antonopoulou, E., Ceppi, I., Sanchez, A., Vernon, E.G., Gamble, A., Myers, K.N., Collis, S.J., Cejka, P., Staples, C.J., 2020. MRNIP is a replication fork protection factor. Sci. Adv. 6, eaba5974. https://doi.org/10.1126/sciadv.aba5974

8.        Sanchez, A., Adam, C., Rauh, F., Duroc, Y., Ranjha, L., Lombard, B., Mu, X., Wintrebert, M., Loew, D., Guarné, A., Gnan, S., Chen, C.-L., Keeney, S., Cejka, P., Guérois, R., Klein, F., Charbonnier, J.-B.,