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Merkel cell polyomavirus recruits MYCL to the EP400 complex to promote oncogenesis
PLoS Pathogens ( IF 6.7 ) Pub Date : 2017-10-13 , DOI: 10.1371/journal.ppat.1006668
Jingwei Cheng , Donglim Esther Park , Christian Berrios , Elizabeth A. White , Reety Arora , Rosa Yoon , Timothy Branigan , Tengfei Xiao , Thomas Westerling , Alexander Federation , Rhamy Zeid , Benjamin Strober , Selene K. Swanson , Laurence Florens , James E. Bradner , Myles Brown , Peter M. Howley , Megha Padi , Michael P. Washburn , James A. DeCaprio

Merkel cell carcinoma (MCC) frequently contains integrated copies of Merkel cell polyomavirus DNA that express a truncated form of Large T antigen (LT) and an intact Small T antigen (ST). While LT binds RB and inactivates its tumor suppressor function, it is less clear how ST contributes to MCC tumorigenesis. Here we show that ST binds specifically to the MYC homolog MYCL (L-MYC) and recruits it to the 15-component EP400 histone acetyltransferase and chromatin remodeling complex. We performed a large-scale immunoprecipitation for ST and identified co-precipitating proteins by mass spectrometry. In addition to protein phosphatase 2A (PP2A) subunits, we identified MYCL and its heterodimeric partner MAX plus the EP400 complex. Immunoprecipitation for MAX and EP400 complex components confirmed their association with ST. We determined that the ST-MYCL-EP400 complex binds together to specific gene promoters and activates their expression by integrating chromatin immunoprecipitation with sequencing (ChIP-seq) and RNA-seq. MYCL and EP400 were required for maintenance of cell viability and cooperated with ST to promote gene expression in MCC cell lines. A genome-wide CRISPR-Cas9 screen confirmed the requirement for MYCL and EP400 in MCPyV-positive MCC cell lines. We demonstrate that ST can activate gene expression in a EP400 and MYCL dependent manner and this activity contributes to cellular transformation and generation of induced pluripotent stem cells.



中文翻译:

默克尔细胞多瘤病毒将MYCL募集到EP400复合物中以促进肿瘤发生

默克尔细胞癌(MCC)通常包含完整的默克尔细胞多瘤病毒DNA拷贝,这些DNA表达截短形式的大T抗原(LT)和完整的小T抗原(ST)。LT结合RB并使其肿瘤抑制功能失活时,尚不清楚ST如何促进MCC肿瘤发生。在这里,我们显示ST特异性结合MYC同源MYCL(L-MYC),并将其募集到15组分EP400组蛋白乙酰转移酶和染色质重塑复合体。我们对ST进行了大规模的免疫沉淀,并通过质谱鉴定了共沉淀蛋白。除了蛋白质磷酸酶2A(PP2A)亚基外,我们还鉴定了MYCL及其异源二聚体伴侣MAX加EP400复合物。MAX和EP400复杂成分的免疫沉淀证实了它们与ST的关联。我们确定ST-MYCL-EP400复合物与特定基因启动子结合在一起,并通过整合染色质免疫沉淀与测序(ChIP-seq)和RNA-seq来激活它们的表达。MYCL和EP400是维持细胞活力所必需的,并与ST协同促进MCC细胞系中的基因表达。全基因组CRISPR-Cas9筛选证实了MCPyV阳性MCC细胞系对MYCL和EP400的需求。我们证明ST可以激活EP400和MYCL依赖方式中的基因表达,这种活动有助于细胞转化和诱导多能干细胞的产生。MYCL和EP400是维持细胞活力所必需的,并与ST协同促进MCC细胞系中的基因表达。全基因组CRISPR-Cas9筛选证实了MCPyV阳性MCC细胞系对MYCL和EP400的需求。我们证明ST可以激活EP400和MYCL依赖方式中的基因表达,这种活动有助于细胞转化和诱导多能干细胞的产生。MYCL和EP400是维持细胞活力所必需的,并与ST协同促进MCC细胞系中的基因表达。全基因组CRISPR-Cas9筛选证实了MCPyV阳性MCC细胞系对MYCL和EP400的需求。我们证明ST可以激活EP400和MYCL依赖方式中的基因表达,这种活动有助于细胞转化和诱导多能干细胞的产生。

更新日期:2017-10-14
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