title: Comparative Proteomics Reveals Strain-Specific β-TrCP Degradation via Rotavirus NSP1 Hijacking a Host Cullin-3-Rbx1 Complex.
abstract: 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.
authors.affiliation: Department of Microbiology and Immunology, 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.|Palo Alto Veterans Institute of Research, VA Palo Alto Health Care System, Palo Alto, California, United States of America.
authors.forename: Siyuan
authors.initials: S
authors.lastname: Ding
authors.affiliation: Department of Microbiology and Immunology, Stanford University School of Medicine, Stanford, California, United States of America.|Baxter Laboratory for Stem Cell Biology, Stanford University School of Medicine, Stanford, California, United States of America.
authors.forename: Nancie
authors.initials: N
authors.lastname: Mooney
authors.affiliation: Department of Medicine, Division of Gastroenterology and Hepatology, Stanford University School of Medicine, Stanford, California, United States of America.|Palo Alto Veterans Institute of Research, VA Palo Alto Health Care System, Palo Alto, California, United States of America.|Institute of Veterinary Medicine, Jiangsu Academy of Agricultural Sciences, Nanjing, China.
authors.forename: Bin
authors.initials: B
authors.lastname: Li
authors.affiliation: Department of Microbiology and Immunology, Stanford University School of Medicine, Stanford, California, United States of America.|Baxter Laboratory for Stem Cell Biology, Stanford University School of Medicine, Stanford, California, United States of America.
authors.forename: Marcus R
authors.initials: MR
authors.lastname: Kelly
authors.affiliation: Department of Microbiology and Immunology, 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.|Palo Alto Veterans Institute of Research, VA Palo Alto Health Care System, Palo Alto, California, United States of America.
authors.forename: Ningguo
authors.initials: N
authors.lastname: Feng
authors.affiliation: Department of Microbiology and Immunology, Stanford University School of Medicine, Stanford, California, United States of America.|Baxter Laboratory for Stem Cell Biology, Stanford University School of Medicine, Stanford, California, United States of America.
authors.forename: Alexander V
authors.initials: AV
authors.lastname: Loktev
authors.affiliation: Department of Microbiology and Immunology, 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.|Palo Alto Veterans Institute of Research, VA Palo Alto Health Care System, Palo Alto, California, United States of America.
authors.forename: Adrish
authors.initials: A
authors.lastname: Sen
authors.affiliation: Department of Veterinary Medicine, University of Maryland, College Park, Maryland, United States of America.
authors.forename: John T
authors.initials: JT
authors.lastname: Patton
authors.affiliation: Department of Microbiology and Immunology, Stanford University School of Medicine, Stanford, California, United States of America.|Baxter Laboratory for Stem Cell Biology, Stanford University School of Medicine, Stanford, California, United States of America.
authors.forename: Peter K
authors.initials: PK
authors.lastname: Jackson
authors.affiliation: Department of Microbiology and Immunology, 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.|Palo Alto Veterans Institute of Research, VA Palo Alto Health Care System, Palo Alto, California, United States of America.
authors.forename: Harry B
authors.initials: HB
authors.lastname: Greenberg
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002352:Carrier Proteins / Q000378:metabolism*; D044842:Cullin Proteins / Q000378:metabolism*; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D057809:HEK293 Cells; D006790:Host-Parasite Interactions / Q000502:physiology*; D006801:Humans; D047468:Immunoprecipitation; D013058:Mass Spectrometry; D040901:Proteomics / Q000379:methods; D060888:Real-Time Polymerase Chain Reaction; D012400:Rotavirus Infections / Q000378:metabolism*; D014162:Transfection; D017361:Viral Nonstructural Proteins / Q000378:metabolism*; D044844:beta-Transducin Repeat-Containing Proteins / Q000378:metabolism*
chemical_list: C503645:CUL3 protein, human; D002352:Carrier Proteins; D044842:Cullin Proteins; C479529:RBX1 protein, human; D017361:Viral Nonstructural Proteins; D044844:beta-Transducin Repeat-Containing Proteins; C104282:nsp1 protein, Rotavirus