Characterization of the protein complex, which is required for the chromatin establishment at the centromere

为了研究着丝粒区域的染色质结构，我们尝试基于高灵敏度质谱结合免疫沉淀来鉴定与CENP-T/CENP-W复合物相关的蛋白。我们鉴定了一种蛋白质Eaf 6，并证实它定位于着丝粒。我们还确定了组蛋白乙酰转移酶作为Eaf 6相互作用蛋白。为了研究Eaf 6的功能，我们建立了一个Eaf 6条件性敲除细胞系，并发现在Eaf 6缺陷细胞中，基于SILAC分析，几个着丝粒蛋白的定位被取消，表明Eaf 6复合物可能是通过Eaf 6复合物的乙酰转移酶活性对着丝粒的染色质结构所必需的。

To examine the chromatin structure in centromere region, we tried to identify proteins that associate with the CENP-T/CENP-W complex based on high sensitive Mass Spectrometry combined with the immunoprecipitation. We identified a protein Eaf6 and we confirmed that this localized to the centromeres. We also identified a histone acethyltransferase as an Eaf6-interacting protein. To examine function of Eaf6, we established an Eaf6-conditional knockout cell line and found that localization of several centromere proteins were abolished in Eaf6-deficient cells based on SILAC analysis, indicating that the Eaf6 complex might be required for chromatin architecture of centromeres through the acethyltransferase activity of the Eaf6 complex.