Checkpoints and Double Strand Breaks in S. Pombe Meiosis

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DESCRIPTION (provided by applicant): Faithful chromosome segregation is essential to the production of viable meiotic products. While the regulation of unperturbed meiotic chromosome segregation is well understood, it is less known what happens when cells attempt meiosis in the presence of unexpected DNA damage. This proposal investigates the response to DNA damage in meiosis, using a fission yeast model system. Fission yeast is a powerful system in the analysis of damage response in the cell cycle, sharing many regulatory genes with humans. Previous studies have shown that the checkpoint system that works in proliferating cells to block the cell cycle in response to DNA damage is not functional in meiosis. Conditions that cause replication fork collapse appear to be compatible with meiotic progression. There is a genetic link between meiotic progression and the response of proliferating cells to alkylation damage that suggest translation synthesis polymerases may play a role in meiosis. These observations suggest that the meiotic response to DNA damage is substantially reprogrammed during differentiation. This is a renewal of a current project that has been funded for 1 year from ARRA (stimulus) funding. The first aim addresses the question of how the damage checkpoint kinase, Chk1, is reprogrammed in meiosis so that it does not respond to damage during S phase. The second aim asks how meiotic cells accommodate collapsing replication forks, which would be lethal during proliferation. The third aim proposes a novel role for trans-lesion synthesis (TLS) polymerases in meiosis. This is based on two observations: first, that the DDK kinase which functions during S phase also regulation meiosis and TLS, and second, that a separation of function allele in the kinase specifically disrupts meiotic divisions and TLS. The long term goal is to understand how the regulation of the damage response during meiS phase is modified to enable later meiotic events. The objective is to use fission yeast to dissect the molecular mechanisms that differ in the response to replication stress and S-phase damage in meiotic cells. The rationale is that knowledge of mechanisms that promote genome stability in meiosis will allow identification of genetic and environmental risk factors that impact human miscarriages and birth defects. The central hypothesis is that conserved activities that normally function to protect the genome are co-opted in meiosis to allow programmed genetic damage. The expected outcomes of this project are the identification and characterization of new molecular pathways. These will include potentially novel factors, likely to be conserved in higher eukaryotes. The positive impact will be a fundamental advance in understanding of the response of differentiating cells to DNA damage and genome stability, and a better understanding of risk factors during meiosis.