Laboratory of Cell Cycle Regulation, Graduate School of Biostudies, Kyoto University, Kyoto, Kyoto 606-8501, Japan.
Department of Stress Response, Radiation Biology Center, Graduate School of Biostudies, Kyoto University, Kyoto, Kyoto 606-8501, Japan.
Nucleic Acids Res. 2021 Oct 11;49(18):10465-10476. doi: 10.1093/nar/gkab767.
Telomere binding protein Stn1 forms the CST (Cdc13/CTC1-STN1-TEN1) complex in budding yeast and mammals. Likewise, fission yeast Stn1 and Ten1 form a complex indispensable for telomere protection. We have previously reported that stn1-1, a high-temperature sensitive mutant, rapidly loses telomere DNA at the restrictive temperature due to frequent failure of replication fork progression at telomeres and subtelomeres, both containing repetitive sequences. It is unclear, however, whether Stn1 is required for maintaining other repetitive DNAs such as ribosomal DNA. In this study, we have demonstrated that stn1-1 cells, even when grown at the permissive temperature, exhibited dynamic rearrangements in the telomere-proximal regions of subtelomere and ribosomal DNA repeats. Furthermore, Rad52 and γH2A accumulation was observed at ribosomal DNA repeats in the stn1-1 mutant. The phenotypes exhibited by the stn1-1 allele were largely suppressed in the absence of Reb1, a replication fork barrier-forming protein, suggesting that Stn1 is involved in the maintenance of the arrested replication forks. Collectively, we propose that Stn1 maintains the stability of repetitive DNAs at subtelomeres and rDNA regions.
端粒结合蛋白 Stn1 在芽殖酵母和哺乳动物中形成 CST(Cdc13/CTC1-STN1-TEN1)复合物。同样,裂殖酵母 Stn1 和 Ten1 形成一个复合物,对于保护端粒是不可或缺的。我们之前曾报道过,高温敏感突变体 stn1-1 在限制温度下由于端粒和亚端粒处复制叉推进的频繁失败,迅速失去端粒 DNA,这两个区域都含有重复序列。然而,目前尚不清楚 Stn1 是否需要维持其他重复 DNA,如核糖体 DNA。在这项研究中,我们已经证明,即使在允许温度下生长,stn1-1 细胞也会在亚端粒和核糖体 DNA 重复序列的端粒近端区域发生动态重排。此外,在 stn1-1 突变体中观察到核糖体 DNA 重复序列处的 Rad52 和 γH2A 积累。在缺乏复制叉障碍形成蛋白 Reb1 的情况下,stn1-1 等位基因的表型得到了很大的抑制,这表明 Stn1 参与维持停滞的复制叉。总的来说,我们提出 Stn1 维持了亚端粒和 rDNA 区域重复 DNA 的稳定性。