Stabilization of G-quadruplex DNA structures in Schizosaccharomyces pombe causes single-strand DNA lesions and impedes DNA replication.
Stabilization of G-quadruplex DNA structures in Schizosaccharomyces pombe causes single-strand DNA lesions and impedes DNA replication.
复制标题
粟酒裂殖酵母中G-四链体DNA结构的稳定性导致单链DNA损伤并阻碍DNA复制。
DOI：
10.1093/nar/gkaa820
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发表时间：
2020-11-04
影响因子：
14.9
通讯作者：
Sabouri N
中科院分区：
文献类型：
--
作者：
Obi I;Rentoft M;Singh V;Jamroskovic J;Chand K;Chorell E;Westerlund F;Sabouri N
G-四链体（G4）结构是稳定的非经典DNA结构，涉及许多细胞途径的调节。我们在这里表明，G4-稳定化合物PhenDC 3引起裂殖酵母细胞的生长缺陷，特别是在同步培养的S期。通过观察单个DNA分子，我们观察到PhenDC 3处理的细胞中新复制的DNA的较短DNA片段，表明PhenDC 3阻碍复制叉进展。此外，一种新的单DNA分子损伤试验显示PhenDC 3处理的细胞中单链DNA损伤增加。此外，染色质免疫沉淀显示，在预测的G4结构的网站的前导链DNA聚合酶的富集，这表明这些结构阻碍DNA复制。我们测试了这些位点的一个子集，并表明它们形成G4结构，它们在体外阻止DNA合成，并且它们可以通过乳腺癌相关的Pif 1家族解旋酶来解决。因此，我们的研究结果表明，G4结构存在于S。粟酒裂殖酵母和稳定/未解决的G4结构是复制机制的障碍。DNA损伤水平的增加可能进一步突出了人类Pif 1解旋酶与家族性乳腺癌以及与未解决的G4结构相关的其他人类疾病的发病之间的关联。
G-quadruplex (G4) structures are stable non-canonical DNA structures that are implicated in the regulation of many cellular pathways. We show here that the G4-stabilizing compound PhenDC3 causes growth defects in Schizosaccharomyces pombe cells, especially during S-phase in synchronized cultures. By visualizing individual DNA molecules, we observed shorter DNA fragments of newly replicated DNA in the PhenDC3-treated cells, suggesting that PhenDC3 impedes replication fork progression. Furthermore, a novel single DNA molecule damage assay revealed increased single-strand DNA lesions in the PhenDC3-treated cells. Moreover, chromatin immunoprecipitation showed enrichment of the leading-strand DNA polymerase at sites of predicted G4 structures, suggesting that these structures impede DNA replication. We tested a subset of these sites and showed that they form G4 structures, that they stall DNA synthesis in vitro and that they can be resolved by the breast cancer-associated Pif1 family helicases. Our results thus suggest that G4 structures occur in S. pombe and that stabilized/unresolved G4 structures are obstacles for the replication machinery. The increased levels of DNA damage might further highlight the association of the human Pif1 helicase with familial breast cancer and the onset of other human diseases connected to unresolved G4 structures.
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DOI：
10.1038/nrg3296
发表时间：
2012-11
期刊：
Nature reviews. Genetics
影响因子：
--
作者：
通讯作者：
--
影响因子：
46.9
作者：
Chambers, Vicki S.;Marsico, Giovanni;Balasubramanian, Shankar
通讯作者：
Balasubramanian, Shankar
影响因子：
3.8
作者：
Bochman ML;Sabouri N;Zakian VA
通讯作者：
Zakian VA
DOI：
10.1016/b978-0-12-387665-2.00006-7
发表时间：
2012
影响因子：
--
作者：
Hegde, Muralidhar L.;Izumi, Tadahide;Mitra, Sankar
通讯作者：
Mitra, Sankar
影响因子：
45.3
作者：
Heinrich, MC;Corless, CL;Fletcher, JA
通讯作者：
Fletcher, JA