Comparative Proteomics Reveals Strain-Specific β-TrCP Degradation via Rotavirus NSP1 Hijacking a Host Cullin-3-Rbx1 Complex

PLoS Pathogens

Harry B. Greenberg¹  Ningguo Feng¹  Nancie Mooney²  Alexander V. Loktev²  Siyuan Ding³  Marcus R. Kelly³  Peter K. Jackson³  John T. Patton⁴  Adrish Sen⁵  Bin Li⁵ 

[1] Baxter Laboratory for Stem Cell Biology, Stanford University School of Medicine, Stanford, California, United States of America;Department of Medicine, Division of Gastroenterology and Hepatology, Stanford University School of Medicine, Stanford, California, United States of America;Department of Microbiology and Immunology, Stanford University School of Medicine, Stanford, California, United States of America;Institute of Veterinary Medicine, Jiangsu Academy of Agricultural Sciences, Nanjing, China;Palo Alto Veterans Institute of Research, VA Palo Alto Health Care System, Palo Alto, California, United States of America

关键词: Small interfering RNAs;    Immunoprecipitation;    Transcription factors;    Ligases;    Cloning;    Host-pathogen interactions;    Proteasomes;    Viral replication;   

DOI  :  10.1371/journal.ppat.1005929

学科分类：生物科学（综合）

来源: Public Library of Science

【 摘 要 】

Rotaviruses (RVs) are the leading cause of severe gastroenteritis in young children, accounting for half a million deaths annually worldwide. RV encodes non-structural protein 1 (NSP1), a well-characterized interferon (IFN) antagonist, which facilitates virus replication by mediating the degradation of host antiviral factors including IRF3 and β-TrCP. Here, we utilized six human and animal RV NSP1s as baits and performed tandem-affinity purification coupled with high-resolution mass spectrometry to comprehensively characterize NSP1-host protein interaction network. Multiple Cullin-RING ubiquitin ligase (CRL) complexes were identified. Importantly, inhibition of cullin-3 (Cul3) or RING-box protein 1 (Rbx1), by siRNA silencing or chemical perturbation, significantly impairs strain-specific NSP1-mediated β-TrCP degradation. Mechanistically, we demonstrate that NSP1 localizes to the Golgi with the host Cul3-Rbx1 CRL complex, which targets β-TrCP and NSP1 for co-destruction at the proteasome. Our study uncovers a novel mechanism that RV employs to promote β-TrCP turnover and provides molecular insights into virus-mediated innate immunity inhibition.

【 授权许可】

CC BY