Cho Yukiko, Ideue Takashi, Nagayama Megumi, Araki Norie, Tani Tokio
Department of Biological Sciences, Graduate School of Science and Technology, Kumamoto University, Kumamoto, Japan.
Department of Tumor Genetics and Biology, Graduate School of Medical Sciences, Kumamoto University, Kumamoto, Japan.
Genes Cells. 2018 Mar;23(3):172-184. doi: 10.1111/gtc.12562. Epub 2018 Jan 31.
Satellite I RNA, a noncoding (nc)RNA transcribed from repetitive regions in human centromeres, binds to Aurora kinase B and forms a ncRNP complex required for chromosome segregation. To examine its function in this process, we purified satellite I ncRNP complex from nuclear extracts prepared from asynchronized or mitotic (M) phase-arrested HeLa cells and then carried out LC/MS to identify proteins bound to satellite I RNA. RBMX (RNA-binding motif protein, X-linked), which was isolated from M phase-arrested cells, was selected for further characterization. We found that RBMX associates with satellite I RNA only during M phase. Knockdown of RBMX induced premature separation of sister chromatid cohesion and abnormal nuclear division. Likewise, knockdown of satellite I RNA also caused premature separation of sister chromatids during M phase. The amounts of RBMX and Sororin, a cohesion regulator, were reduced in satellite I RNA-depleted cells. These results suggest that satellite I RNA plays a role in stabilizing RBMX and Sororin in the ncRNP complex to maintain proper sister chromatid cohesion.
卫星I RNA是一种从人类着丝粒重复区域转录而来的非编码(nc)RNA,它与极光激酶B结合,形成染色体分离所需的ncRNP复合物。为了研究其在此过程中的功能,我们从处于非同步或有丝分裂(M)期阻滞的HeLa细胞制备的核提取物中纯化卫星I ncRNP复合物,然后进行液相色谱/质谱分析以鉴定与卫星I RNA结合的蛋白质。从M期阻滞细胞中分离出的RBMX(RNA结合基序蛋白,X连锁)被选作进一步表征。我们发现RBMX仅在M期与卫星I RNA结合。敲低RBMX会导致姐妹染色单体黏连过早分离和异常核分裂。同样,敲低卫星I RNA也会在M期导致姐妹染色单体过早分离。在卫星I RNA缺失的细胞中,RBMX和黏连调节因子Sororin的量减少。这些结果表明,卫星I RNA在ncRNP复合物中对稳定RBMX和Sororin起作用,以维持适当的姐妹染色单体黏连。