Molecular mechanism of EPC1-PHF1 fusion protein (CUT and RUN-seq Homo Sapiens)

In this study, we characterize the fusion protein produced by the EPC1-PHF1 translocation in Low Grade Endometrial Stromal Sarcoma (LG-ESS) and Ossifying FibroMyxoid Tumors (OFMT). We express the fusion protein and necessary controls in K562 Cells. The fusion protein assembles a mega-complex harboring both NuA4/TIP60 and PRC2 subunits and enzymatic activities and leads to mislocalization of chromatin marks in the genome, linked to aberrant gene expression. CUT and RUN-sequencing (anti-HA) in K562 cells expressing HA tagged EPC1-PHF1 fusion protein or tag only.

本研究针对低级别子宫内膜间质肉瘤（Low Grade Endometrial Stromal Sarcoma, LG-ESS）与钙化性纤维黏液瘤（Ossifying FibroMyxoid Tumors, OFMT）中由EPC1-PHF1易位产生的融合蛋白进行了系统表征。我们在K562细胞中表达该融合蛋白及必要的对照蛋白。该融合蛋白可组装形成巨型复合物，该复合物同时包含NuA4/TIP60与PRC2亚基并具备相应酶活性，进而导致基因组中染色质标记的异位分布，这一改变与异常基因表达密切相关。本研究在表达经HA标签标记的EPC1-PHF1融合蛋白或仅表达HA标签的K562细胞中，开展了基于抗HA抗体的CUT&RUN测序（CUT and RUN-sequencing）实验。