title: Genome-Wide Screen Reveals Valosin-Containing Protein Requirement for Coronavirus Exit from Endosomes.
abstract: Coronaviruses are RNA viruses with a large zoonotic reservoir and propensity for host switching, representing a real threat for public health, as evidenced by severe acute respiratory syndrome (SARS) and the emerging Middle East respiratory syndrome (MERS). Cellular factors required for their replication are poorly understood. Using genome-wide small interfering RNA (siRNA) screening, we identified 83 novel genes supporting infectious bronchitis virus (IBV) replication in human cells. Thirty of these hits can be placed in a network of interactions with viral proteins and are involved in RNA splicing, membrane trafficking, and ubiquitin conjugation. In addition, our screen reveals an unexpected role for valosin-containing protein (VCP/p97) in early steps of infection. Loss of VCP inhibits a previously uncharacterized degradation of the nucleocapsid N protein. This inhibition derives from virus accumulation in early endosomes, suggesting a role for VCP in the maturation of virus-loaded endosomes. The several host factors identified in this study may provide avenues for targeted therapeutics.
  
  
  OBJECTIVE
  Coronaviruses are RNA viruses representing a real threat for public health, as evidenced by SARS and the emerging MERS. However, cellular factors required for their replication are poorly understood. Using genome-wide siRNA screening, we identified novel genes supporting infectious bronchitis virus (IBV) replication in human cells. The several host factors identified in this study may provide directions for future research on targeted therapeutics.
authors: Institute of Molecular and Cell Biology/A*STAR, Singapore, Singapore Department of Biochemistry, Yong Loo Lin School of Medicine, National University of Singapore, Singapore, Singapore.
authors: Hui Hui
authors: HH
authors: Wong
authors: Institute of Molecular and Cell Biology/A*STAR, Singapore, Singapore.
authors: Pankaj
authors: P
authors: Kumar
authors: Institute of Molecular and Cell Biology/A*STAR, Singapore, Singapore.
authors: Felicia Pei Ling
authors: FP
authors: Tay
authors: Institute of Molecular and Cell Biology/A*STAR, Singapore, Singapore.
authors: Dimitri
authors: D
authors: Moreau
authors: Institute of Molecular and Cell Biology/A*STAR, Singapore, Singapore School of Biological Sciences, Nanyang Technological University, Singapore, Singapore dxliu@ntu.edu.sg fbard@imcb.a-star.edu.sg.
authors: Ding Xiang
authors: DX
authors: Liu
authors: Institute of Molecular and Cell Biology/A*STAR, Singapore, Singapore Department of Biochemistry, Yong Loo Lin School of Medicine, National University of Singapore, Singapore, Singapore dxliu@ntu.edu.sg fbard@imcb.a-star.edu.sg.
authors: Frédéric
authors: F
authors: Bard
mesh_terms: D000251:Adenosine Triphosphatases / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D015153:Blotting, Western; D018797:Cell Cycle Proteins / Q000235:genetics / Q000378:metabolism*; D002460:Cell Line; D002522:Chlorocebus aethiops; D018352:Coronavirus Infections / Q000635:transmission*; D011992:Endosomes / Q000821:virology*; D005455:Fluorescent Antibody Technique; D055106:Genome-Wide Association Study; D006801:Humans; D001351:Infectious bronchitis virus / Q000502:physiology*; D058977:Molecular Sequence Annotation; D034741:RNA, Small Interfering / Q000235:genetics; D000074405:Valosin Containing Protein; D014709:Vero Cells; D057074:Virus Release / Q000502:physiology*
chemical_list: D018797:Cell Cycle Proteins; D034741:RNA, Small Interfering; D000251:Adenosine Triphosphatases; C000614178:VCP protein, human; D000074405:Valosin Containing Protein