DNA double-strand break (DSB) repair is critical to prevent persistent DNA damage, counteract genome instability and suppress tumor development. By re-joining broken DNA ends, DSB repair has also the potential to produce chromosome translocations and complex rearrangements. In this project, we will investigate the mechanism and regulation of the earliest step of DSB repair, DNA end-processing and its impact on DSB repair choice, particularly microhomology-mediated end-joining (MMEJ). We hypothesize that persistent, short ssDNA overhangs is the aberrant DNA repair intermediate involved in MMEJ, the pathogenic form of DSB repair responsible for a significant fraction of tumorigenic chromosome translocations. First, we will delineate the conditions that favor the generation of these intennediates in cell-free extracts derived from Xenopus. We will evaluate the contribution of the three resection pathways: CtlP, EX01 and DN/^2. Next, we will assess the impact of cell cycle (CDK) and DNA damage (PIKK) protein kinases on these pathways and on DNA processing. Finally, we will evaluate how DNA resection influences MMEJ in human breast tumor cells with fully sequenced genomes and documented chromosome translocation landscapes. The biochemical findings gained from this project will inform experiments in all other projects of the program. The proposed experiments should provide an unprecedented level of understanding of the mechanism and regulation of resection of ligatable DNA ends as well as damaged DNA ends or DNA ends harboring adducts, such as those generated following chemo- or radiotherapy. In turn this will guide future studies aiming at decreasing the impact of chromosome translocations and improving the efficiency of cancer therapies based on the generation of DNA DSBs.
Aberrant chromosome rearrangements, particulariy chromosome translocations, are implicated in the development of human cancers. Translocations often arise from a pathogenic form of DNA repair that generates short single-strand DNA intermediates with microhomologies (MH). The goal of this project is to determine how resection and the DNA repair protein, CtlP, participate in generating persistent, toxic repair intermediates with MH leading to chromosome translocations causing tumor development.
|Yu, Tai-Yuan; Kimble, Michael T; Symington, Lorraine S (2018) Sae2 antagonizes Rad9 accumulation at DNA double-strand breaks to attenuate checkpoint signaling and facilitate end resection. Proc Natl Acad Sci U S A 115:E11961-E11969|
|Oh, Julyun; Lee, So Jung; Rothstein, Rodney et al. (2018) Xrs2 and Tel1 Independently Contribute to MR-Mediated DNA Tethering and Replisome Stability. Cell Rep 25:1681-1692.e4|
|Billing, David; Horiguchi, Michiko; Wu-Baer, Foon et al. (2018) The BRCT Domains of the BRCA1 and BARD1 Tumor Suppressors Differentially Regulate Homology-Directed Repair and Stalled Fork Protection. Mol Cell 72:127-139.e8|
|Schrank, Benjamin R; Aparicio, Tomas; Li, Yinyin et al. (2018) Nuclear ARP2/3 drives DNA break clustering for homology-directed repair. Nature 559:61-66|
|Gnügge, Robert; Oh, Julyun; Symington, Lorraine S (2018) Processing of DNA Double-Strand Breaks in Yeast. Methods Enzymol 600:1-24|
|Crowe, Jennifer L; Shao, Zhengping; Wang, Xiaobin S et al. (2018) Kinase-dependent structural role of DNA-PKcs during immunoglobulin class switch recombination. Proc Natl Acad Sci U S A 115:8615-8620|
|Gnügge, Robert; Symington, Lorraine S (2017) Keeping it real: MRX-Sae2 clipping of natural substrates. Genes Dev 31:2311-2312|
|Liu, Xiangyu; Shao, Zhengping; Jiang, Wenxia et al. (2017) PAXX promotes KU accumulation at DNA breaks and is essential for end-joining in XLF-deficient mice. Nat Commun 8:13816|
|Kato, Niyo; Kawasoe, Yoshitaka; Williams, Hannah et al. (2017) Sensing and Processing of DNA Interstrand Crosslinks by the Mismatch Repair Pathway. Cell Rep 21:1375-1385|
|Aparicio, Tomas; Baer, Richard; Gottesman, Max et al. (2016) MRN, CtIP, and BRCA1 mediate repair of topoisomerase II-DNA adducts. J Cell Biol 212:399-408|
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