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).

以前，我发现RSF复合物会与CENP-A瞬时相互作用，并在CENP-A掺入中心粒中作用。因此，我想确定1）RSF复合物如何靶向CentRomere，2）是帮助RSF复合物在CENP-A染色质的稳定保留和3）3）RSF复合物中如何参与动力学组合体中涉及RSF复合物的协同因素。为了解决这些问题，我对DT40细胞的抗FLAG染色质免疫沉淀稳定地表达了FLAG-RSF-I，发现与RSF相关的DNA沿基因组分布，但是特定的尖峰与CENP-A相关DNA的峰重叠。此外，我发现RSF复合物主要与事实复合物相互作用，这是另一种染色质重塑复合物，我们也在Centromere发现，发现与CENP-A相互作用。我证明了通过FACTP80（SSRP1）敲除细胞中的RSF1量减少。

项目成果

The ABCs of CENPs

• DOI: 10.1007/s00412-011-0330-0
• 发表时间: 2011-10-01
• 期刊: CHROMOSOMA
• 影响因子: 1.6
• 作者: Perpelescu, Marinela;Fukagawa, Tatsuo
• 通讯作者: Fukagawa, Tatsuo

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