{"title":"SMARCA2 和 SMARCA4 参与 DNA 损伤修复","authors":"Lily Yu, Duo Wu","doi":"10.31083/j.fbl2907262","DOIUrl":null,"url":null,"abstract":"<p><strong>Background: </strong>The switching/sucrose non-fermentable (SWI/SNF) Related, Matrix Associated, Actin Dependent Regulator Of Chromatin, Subfamily A (SMARCA) member 2 and member 4 (SMARCA2/4) are paralogs and act as the key enzymatic subunits in the SWI/SNF complex for chromatin remodeling. However, the role of SMARCA2/4 in DNA damage response remains unclear.</p><p><strong>Methods: </strong>Laser microirradiation assays were performed to examine the key domains of SMARCA2/4 for the relocation of the SWI/SNF complex to DNA lesions. To examine the key factors that mediate the recruitment of SMARCA2/4, the relocation of SMARCA2/4 to DNA lesions was examined in HeLa cells treated with inhibitors of Ataxia-telangiectasia-mutated (ATM), Ataxia telangiectasia and Rad3-related protein (ATR), CREB-binding protein (CBP) and its homologue p300 (p300/CBP), or Poly (ADP-ribose) polymerase (PARP) 1/2 as well as in H2AX-deficient HeLa cells. Moreover, by concomitantly suppressing SMARCA2/4 with the small molecule inhibitor FHD286 or Compound 14, the function of SMARCA2/4 in Radiation sensitive 51 (RAD51) foci formation and homologous recombination repair was examined. Finally, using a colony formation assay, the synergistic effect of PARP inhibitors and SMARCA2/4 inhibitors on the suppression of tumor cell growth was examined.</p><p><strong>Results: </strong>We show that SMARCA2/4 relocate to DNA lesions in response to DNA damage, which requires their ATPase activities. Moreover, these ATPase activities are also required for the relocation of other subunits in the SWI/SNF complex to DNA lesions. Interestingly, the relocation of SMARCA2/4 is independent of γH2AX, ATM, ATR, p300/CBP, or PARP1/2, indicating that it may directly recognize DNA lesions as a DNA damage sensor. Lacking SMARCA2/4 prolongs the retention of γH2AX, Ring Finger Protein 8 (RNF8) and Breast cancer susceptibility gene 1 (BRCA1) at DNA lesions and impairs RAD51-dependent homologous recombination repair. Furthermore, the treatment of an SMARCA2/4 inhibitor sensitizes tumor cells to PARP inhibitor treatment.</p><p><strong>Conclusions: </strong>This study reveals SMARCA2/4 as a DNA damage repair factor for double-strand break repair.</p>","PeriodicalId":73069,"journal":{"name":"Frontiers in bioscience (Landmark edition)","volume":null,"pages":null},"PeriodicalIF":3.3000,"publicationDate":"2024-07-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"SMARCA2 and SMARCA4 Participate in DNA Damage Repair.\",\"authors\":\"Lily Yu, Duo Wu\",\"doi\":\"10.31083/j.fbl2907262\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><strong>Background: </strong>The switching/sucrose non-fermentable (SWI/SNF) Related, Matrix Associated, Actin Dependent Regulator Of Chromatin, Subfamily A (SMARCA) member 2 and member 4 (SMARCA2/4) are paralogs and act as the key enzymatic subunits in the SWI/SNF complex for chromatin remodeling. However, the role of SMARCA2/4 in DNA damage response remains unclear.</p><p><strong>Methods: </strong>Laser microirradiation assays were performed to examine the key domains of SMARCA2/4 for the relocation of the SWI/SNF complex to DNA lesions. To examine the key factors that mediate the recruitment of SMARCA2/4, the relocation of SMARCA2/4 to DNA lesions was examined in HeLa cells treated with inhibitors of Ataxia-telangiectasia-mutated (ATM), Ataxia telangiectasia and Rad3-related protein (ATR), CREB-binding protein (CBP) and its homologue p300 (p300/CBP), or Poly (ADP-ribose) polymerase (PARP) 1/2 as well as in H2AX-deficient HeLa cells. Moreover, by concomitantly suppressing SMARCA2/4 with the small molecule inhibitor FHD286 or Compound 14, the function of SMARCA2/4 in Radiation sensitive 51 (RAD51) foci formation and homologous recombination repair was examined. Finally, using a colony formation assay, the synergistic effect of PARP inhibitors and SMARCA2/4 inhibitors on the suppression of tumor cell growth was examined.</p><p><strong>Results: </strong>We show that SMARCA2/4 relocate to DNA lesions in response to DNA damage, which requires their ATPase activities. Moreover, these ATPase activities are also required for the relocation of other subunits in the SWI/SNF complex to DNA lesions. Interestingly, the relocation of SMARCA2/4 is independent of γH2AX, ATM, ATR, p300/CBP, or PARP1/2, indicating that it may directly recognize DNA lesions as a DNA damage sensor. Lacking SMARCA2/4 prolongs the retention of γH2AX, Ring Finger Protein 8 (RNF8) and Breast cancer susceptibility gene 1 (BRCA1) at DNA lesions and impairs RAD51-dependent homologous recombination repair. Furthermore, the treatment of an SMARCA2/4 inhibitor sensitizes tumor cells to PARP inhibitor treatment.</p><p><strong>Conclusions: </strong>This study reveals SMARCA2/4 as a DNA damage repair factor for double-strand break repair.</p>\",\"PeriodicalId\":73069,\"journal\":{\"name\":\"Frontiers in bioscience (Landmark edition)\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":3.3000,\"publicationDate\":\"2024-07-23\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Frontiers in bioscience (Landmark edition)\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://doi.org/10.31083/j.fbl2907262\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q2\",\"JCRName\":\"BIOCHEMISTRY & MOLECULAR BIOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Frontiers in bioscience (Landmark edition)","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.31083/j.fbl2907262","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"BIOCHEMISTRY & MOLECULAR BIOLOGY","Score":null,"Total":0}
引用次数: 0
摘要
背景：开关/蔗糖不发酵（SWI/SNF）相关、基质相关、肌动蛋白依赖的染色质调节器亚家族 A（SMARCA）成员 2 和成员 4（SMARCA2/4）是对映体，是 SWI/SNF 复合物中染色质重塑的关键酶亚基。然而，SMARCA2/4在DNA损伤反应中的作用仍不清楚：方法：通过激光微辐照实验来研究 SMARCA2/4 在将 SWI/SNF 复合物迁移到 DNA 损伤点时的关键结构域。为了研究介导SMARCA2/4招募的关键因素，研究人员在用共济失调-毛细血管扩张症突变（ATM）抑制剂处理的HeLa细胞中检测了SMARCA2/4向DNA损伤部位的迁移、ATR）、CREB 结合蛋白（CBP）及其同源物 p300（p300/CBP）或聚（ADP-核糖）聚合酶（PARP）1/2 的抑制剂处理的 HeLa 细胞以及 H2AX 缺失的 HeLa 细胞中，研究了 SMARCA2/4 在 DNA 病变处的迁移情况。此外，通过用小分子抑制剂 FHD286 或化合物 14 同时抑制 SMARCA2/4，研究了 SMARCA2/4 在辐射敏感 51（RAD51）病灶形成和同源重组修复中的功能。最后，利用集落形成试验，研究了 PARP 抑制剂和 SMARCA2/4 抑制剂对抑制肿瘤细胞生长的协同作用：结果：我们发现，SMARCA2/4在DNA损伤时会迁移到DNA病变部位，这需要它们的ATP酶活性。此外，SWI/SNF 复合物中的其他亚基迁移到 DNA 损伤处也需要这些 ATPase 活性。有趣的是，SMARCA2/4 的迁移与 γH2AX、ATM、ATR、p300/CBP 或 PARP1/2 无关，这表明它可能作为 DNA 损伤传感器直接识别 DNA 病变。缺乏 SMARCA2/4，γH2AX、环指蛋白 8（RNF8）和乳腺癌易感基因 1（BRCA1）在 DNA 病变处的滞留时间就会延长，并损害 RAD51 依赖性同源重组修复。此外，SMARCA2/4抑制剂可使肿瘤细胞对PARP抑制剂治疗敏感：本研究揭示了 SMARCA2/4 是一种用于双链断裂修复的 DNA 损伤修复因子。
SMARCA2 and SMARCA4 Participate in DNA Damage Repair.
Background: The switching/sucrose non-fermentable (SWI/SNF) Related, Matrix Associated, Actin Dependent Regulator Of Chromatin, Subfamily A (SMARCA) member 2 and member 4 (SMARCA2/4) are paralogs and act as the key enzymatic subunits in the SWI/SNF complex for chromatin remodeling. However, the role of SMARCA2/4 in DNA damage response remains unclear.
Methods: Laser microirradiation assays were performed to examine the key domains of SMARCA2/4 for the relocation of the SWI/SNF complex to DNA lesions. To examine the key factors that mediate the recruitment of SMARCA2/4, the relocation of SMARCA2/4 to DNA lesions was examined in HeLa cells treated with inhibitors of Ataxia-telangiectasia-mutated (ATM), Ataxia telangiectasia and Rad3-related protein (ATR), CREB-binding protein (CBP) and its homologue p300 (p300/CBP), or Poly (ADP-ribose) polymerase (PARP) 1/2 as well as in H2AX-deficient HeLa cells. Moreover, by concomitantly suppressing SMARCA2/4 with the small molecule inhibitor FHD286 or Compound 14, the function of SMARCA2/4 in Radiation sensitive 51 (RAD51) foci formation and homologous recombination repair was examined. Finally, using a colony formation assay, the synergistic effect of PARP inhibitors and SMARCA2/4 inhibitors on the suppression of tumor cell growth was examined.
Results: We show that SMARCA2/4 relocate to DNA lesions in response to DNA damage, which requires their ATPase activities. Moreover, these ATPase activities are also required for the relocation of other subunits in the SWI/SNF complex to DNA lesions. Interestingly, the relocation of SMARCA2/4 is independent of γH2AX, ATM, ATR, p300/CBP, or PARP1/2, indicating that it may directly recognize DNA lesions as a DNA damage sensor. Lacking SMARCA2/4 prolongs the retention of γH2AX, Ring Finger Protein 8 (RNF8) and Breast cancer susceptibility gene 1 (BRCA1) at DNA lesions and impairs RAD51-dependent homologous recombination repair. Furthermore, the treatment of an SMARCA2/4 inhibitor sensitizes tumor cells to PARP inhibitor treatment.
Conclusions: This study reveals SMARCA2/4 as a DNA damage repair factor for double-strand break repair.
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