Functional role of the RSF complex in establishment of centromere-specific chromatin

RSF 复合物在着丝粒特异性染色质建立中的功能作用

• 批准号: 22770177
• 负责人: PERPELESCU Marinela
• 金额: $ 2.75万
• 依托单位: National Institute of Genetics
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Young Scientists (B)
• 财政年份: 2010
• 资助国家: 日本
• 起止时间: 2010 至 2011
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-22770177/
• 关键词: セントロメア; クロマチン; CENP-A; RSF; 細胞周期; DT40細胞; 染色体分配; 免疫沈降

Previously, I found that the RSF complex transiently interacts with CENP-A and acts in CENP-A incorporation into centromeres. Therefore, I wanted to determine 1) how the RSF complex targets the centromere, 2) which are the synergic factors that help the RSF complex in the stabile retention of CENP-A chromatin and 3) how the RSF complex is involved in kinetochore assembly. To address these questions, I performed anti-FLAG chromatin immunoprecipitation of DT40 cells stably expressing FLAG-Rsf-I and found that RSF-associated DNA distributes along the genome, but a specific sharp peak overlaps the peak of the CENP-A-associated DNA. In addition, I found that the RSF complex mainly interacts with the FACT complex, which is another chromatin remodeling complex that we also spotted at centromere and found that interacts with CENP-A. I demonstrated that the Rsf1 amount was reduced in cells with knockout of FACTp80(SSRP1).

此前，我发现 RSF 复合物与 CENP-A 短暂相互作用，并在 CENP-A 掺入着丝粒中发挥作用。因此，我想确定 1) RSF 复合物如何靶向着丝粒，2) 哪些协同因素有助于 RSF 复合物稳定保留 CENP-A 染色质，3) RSF 复合物如何参与着丝粒组装。为了解决这些问题，我对稳定表达 FLAG-Rsf-I 的 DT40 细胞进行了抗 FLAG 染色质免疫沉淀，发现 RSF 相关 DNA 沿基因组分布，但有一个特定的尖峰与 CENP-A 相关 DNA 的峰重叠。此外，我发现 RSF 复合物主要与 FACT 复合物相互作用，FACT 复合物是我们也在着丝粒处发现的另一种染色质重塑复合物，并发现它与 CENP-A 相互作用。我证明敲除 FACTp80(SSRP1) 的细胞中 Rsf1 的量减少。

项目成果

CENTROMERE CHROMATIN REMODELING-A COOPERATIVE WORK

着丝粒染色质重塑——一项合作工作

• 发表时间: 2012
• 作者: Liu J;Chen CY;Shiomi D;Niki H;Margolin W.;Perpelescu Marinela
• 通讯作者: Perpelescu Marinela