Merkel cell polyomavirus recruits MYCL to the EP400 complex to promote oncogenesis

PLoS Pathogens

Alexander Federation¹  Tengfei Xiao¹  Peter M. Howley²  Laurence Florens³  Jingwei Cheng³  Benjamin Strober³  Rosa Yoon³  Megha Padi³  Myles Brown³  Elizabeth A. White³  Rhamy Zeid³  Michael P. Washburn⁴  Donglim Esther Park⁴  James A. DeCaprio⁵  Christian Berrios⁵  Timothy Branigan⁶  Thomas Westerling⁶  Selene K. Swanson⁷  James E. Bradner⁷  Reety Arora⁷ 

[1]Boston, Massachusetts, United States of America

[2]Center for Functional Cancer Epigenetics, Dana-Farber Cancer Institute, Boston, Massachusetts, United States of America

[3]Department of Medical Oncology, Dana-Farber Cancer Institute, Boston, Massachusetts, United States of America

[4]Department of Medicine, Brigham and Women’s Hospital, Boston, Massachusetts, United States of America

[5]Department of Medicine, Harvard Medical School, Boston, Massachusetts, United States of America

[6]Department of Microbiology and Immunobiology, Harvard Medical School

[7]Graduate School of Arts and Sciences, Harvard University, Boston, Massachusetts, United States of America

关键词: Immunoprecipitation;    Gene expression;    Carcinogenesis;    CRISPR;    Chromatin;    Induced pluripotent stem cells;    Library screening;    Gene ontologies;   

DOI  :  10.1371/journal.ppat.1006668

学科分类：生物科学（综合）

来源: Public Library of Science

【 摘 要 】

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.

【 授权许可】

CC BY