Huang Pei-Ching, Hong Soogil, Alnaser Hasan F, Mimitou Eleni P, Kim Keun P, Murakami Hajime, Keeney Scott
Weill Cornell Graduate School of Medical Sciences, Cornell University, New York, NY, 10021, USA.
Molecular Biology Program, Memorial Sloan Kettering Cancer Center, New York, NY, 10065, USA.
EMBO J. 2025 Jan;44(1):200-224. doi: 10.1038/s44318-024-00318-8. Epub 2024 Nov 29.
DNA double-strand breaks (DSBs) are nucleolytically processed to generate single-stranded DNA for homologous recombination. In Saccharomyces cerevisiae meiosis, this resection involves nicking by the Mre11-Rad50-Xrs2 complex (MRX), then exonucleolytic digestion by Exo1. Chromatin remodeling at meiotic DSBs is thought necessary for resection, but the remodeling enzyme was unknown. Here we show that the SWI/SNF-like ATPase Fun30 plays a major, nonredundant role in meiotic resection. A fun30 mutation shortened resection tracts almost as severely as an exo1-nd (nuclease-dead) mutation, and resection was further shortened in a fun30 exo1-nd double mutant. Fun30 associates with chromatin in response to DSBs, and the constitutive positioning of nucleosomes governs resection endpoint locations in the absence of Fun30. We infer that Fun30 promotes both the MRX- and Exo1-dependent steps in resection, possibly by removing nucleosomes from broken chromatids. Moreover, the extremely short resection in fun30 exo1-nd double mutants is accompanied by compromised interhomolog recombination bias, leading to defects in recombination and chromosome segregation. Thus, this study also provides insight about the minimal resection lengths needed for robust recombination.
DNA双链断裂(DSBs)经核酸酶处理后生成单链DNA用于同源重组。在酿酒酵母减数分裂过程中,这种切除涉及Mre11-Rad50-Xrs2复合物(MRX)的切口作用,然后由Exo1进行核酸外切消化。减数分裂DSBs处的染色质重塑被认为是切除过程所必需的,但重塑酶尚不清楚。在这里,我们表明SWI/SNF样ATP酶Fun30在减数分裂切除中起主要的、非冗余的作用。fun30突变导致切除片段缩短的程度几乎与exo1-nd(核酸酶失活)突变一样严重,而在fun30 exo1-nd双突变体中切除进一步缩短。Fun30响应DSBs与染色质结合,并且在没有Fun30的情况下,核小体的组成性定位决定了切除终点的位置。我们推断Fun30可能通过从断裂的染色单体上去除核小体来促进切除过程中依赖MRX和Exo1的步骤。此外,fun30 exo1-nd双突变体中极短的切除伴随着同源重组偏向性受损,导致重组和染色体分离缺陷。因此,本研究还提供了关于强大重组所需的最小切除长度的见解。