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.
A substantial fraction of birth defects result from chromosomal defects in meiosis, the process that produces eggs and sperm. This project uses genetics and cell biology in simple yeast to study how chromosomes in meiosis are protected from DNA damage that may contribute to meiotic defects. The goal is to identify conserved proteins that protect meiotic cells from defects that could contribute to birth defects.
|Escorcia, Wilber; Forsburg, Susan L (2017) Destabilization of the replication fork protection complex disrupts meiotic chromosome segregation. Mol Biol Cell 28:2978-2997|
|Ranatunga, Nimna S; Forsburg, Susan L (2016) Characterization of a Novel MMS-Sensitive Allele of Schizosaccharomyces pombe mcm4. G3 (Bethesda) 6:3049-3063|
|Sabatinos, Sarah A; Ranatunga, Nimna S; Yuan, Ji-Ping et al. (2015) Replication stress in early S phase generates apparent micronuclei and chromosome rearrangement in fission yeast. Mol Biol Cell 26:3439-50|
|Sabatinos, Sarah A; Forsburg, Susan L (2015) Managing Single-Stranded DNA during Replication Stress in Fission Yeast. Biomolecules 5:2123-39|
|Ding, Lin; Forsburg, Susan L (2014) Essential domains of Schizosaccharomyces pombe Rad8 required for DNA damage response. G3 (Bethesda) 4:1373-84|
|Mastro, Tara L; Forsburg, Susan L (2014) Increased meiotic crossovers and reduced genome stability in absence of Schizosaccharomyces pombe Rad16 (XPF). Genetics 198:1457-72|
|Ding, Lin; Laor, Dana; Weisman, Ronit et al. (2014) Rapid regulation of nuclear proteins by rapamycin-induced translocation in fission yeast. Yeast 31:253-64|
|Sabatinos, Sarah A; Mastro, Tara L; Green, Marc D et al. (2013) A mammalian-like DNA damage response of fission yeast to nucleoside analogs. Genetics 193:143-57|
|Le, Anh-Huy; Mastro, Tara L; Forsburg, Susan L (2013) The C-terminus of S. pombe DDK subunit Dfp1 is required for meiosis-specific transcription and cohesin cleavage. Biol Open 2:728-38|
|Dolan, William P; Le, Anh-Huy; Schmidt, Henning et al. (2010) Fission yeast Hsk1 (Cdc7) kinase is required after replication initiation for induced mutagenesis and proper response to DNA alkylation damage. Genetics 185:39-53|