Comprehensive analysis of the ICEN (Interphase Centromere Complex) components enriched in the CENP-A chromatin of human cells
Comprehensive analysis of the ICEN (Interphase Centromere Complex) components enriched in the CENP-A chromatin of human cells
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DOI：
10.1111/j.1365-2443.2006.00969.x
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发表时间：
2006-06-01
期刊：
影响因子：
2.1
通讯作者：
Yoda, Kinya
中科院分区：
文献类型：
--
作者：
Izuta, Hiroshi;Ikeno, Masashi;Yoda, Kinya
着丝粒是姐妹染色单体正确分离所必需的染色质结构，该区域的缺陷通常导致非整倍性和癌症。我们以前曾报道过从HeLa细胞中纯化间期着丝粒复合物（ICEN），并通过蛋白质组学分析证明了40种蛋白质（ICEN 1 -40）的存在，沿着CENP-A、-B、-C、-H和hMis 6。在这里，我们报告了7个ICEN组件与未知功能的分析。EGFP标记的ICEN 22、24、32、33、36、37和39的着丝粒定位在p53细胞中观察到。这些蛋白质的每一个短RNA干扰产生的异常中期细胞携带错位的染色体，也产生含有非整倍体染色体的细胞，这意味着这些ICEN蛋白参与动粒功能。有趣的是，在ICEN 22、32、33、37或39 siRNA转染的细胞中，CENP-H和hMis 6信号从含有错配染色体的异常有丝分裂细胞中的所有着丝粒消失。这些结果表明，ICEN复合物的7个组分主要定位于着丝粒，并且可能通过或不通过将CENP-H和hMis 6加载到着丝粒上而为动粒功能所需。
The centromere is a chromatin structure essential for correct segregation of sister chromatids, and defects in this region often lead to aneuploidy and cancer. We have previously reported purification of the interphase centromere complex (ICEN) from HeLa cells, and have demonstrated the presence of 40 proteins (ICEN1-40), along with CENP-A, -B, -C, -H and hMis6, by proteomic analysis. Here we report analysis of seven ICEN components with unknown function. Centromere localization of EGFP-tagged ICEN22, 24, 32, 33, 36, 37 and 39 was observed in transformant cells. Depletion of each of these proteins by short RNA interference produced abnormal metaphase cells carrying misaligned chromosomes and also produced cells containing aneuploid chromosomes, implying that these ICEN proteins take part in kinetochore functions. Interestingly, in the ICEN22, 32, 33, 37 or 39 siRNA-transfected cells, CENP-H and hMis6 signals disappeared from all the centromeres in abnormal mitotic cells containing misaligned chromosomes. These results suggest that the seven components of the ICEN complex are predominantly localized at the centromeres and are required for kinetochore function perhaps through or not through loading of CENP-H and hMis6 onto the centromere.