title: Antagonistic effects of cellular poly(C) binding proteins on vesicular stomatitis virus gene expression.
abstract: Immunoprecipitation and subsequent mass spectrometry analysis of the cellular proteins from cells expressing the vesicular stomatitis virus (VSV) P protein identified the poly(C) binding protein 2 (PCBP2) as one of the P protein-interacting proteins. To investigate the role of PCBP2 in the viral life cycle, we examined the effects of depletion or overexpression of this protein on VSV growth. Small interfering RNA-mediated silencing of PCBP2 promoted VSV replication. Conversely, overexpression of PCBP2 in transfected cells suppressed VSV growth. Further studies revealed that PCBP2 negatively regulates overall viral mRNA accumulation and subsequent genome replication. Coimmunoprecipitation and immunofluorescence microscopic studies showed that PCBP2 interacts and colocalizes with VSV P protein in virus-infected cells. The P-PCBP2 interaction did not result in reduced levels of protein complex formation with the viral N and L proteins, nor did it induce degradation of the P protein. In addition, PCBP1, another member of the poly(C) binding protein family with homology to PCBP2, was also found to interact with the P protein and inhibit the viral mRNA synthesis at the level of primary transcription without affecting secondary transcription or genome replication. The inhibitory effects of PCBP1 on VSV replication were less pronounced than those of PCBP2. Overall, the results presented here suggest that cellular PCBP2 and PCBP1 antagonize VSV growth by affecting viral gene expression and highlight the importance of these two cellular proteins in restricting virus infections.
authors.affiliation: 109 Morrison Life Science Research Center, 4240 Fair Street, East Campus, University of Nebraska-Lincoln, Lincoln, NE 68583-0900, USA.
authors.forename: Phat X
authors.initials: PX
authors.lastname: Dinh
authors.forename: Lalit K
authors.initials: LK
authors.lastname: Beura
authors.forename: Debasis
authors.initials: D
authors.lastname: Panda
authors.forename: Anshuman
authors.initials: A
authors.lastname: Das
authors.forename: Asit K
authors.initials: AK
authors.lastname: Pattnaik
mesh_terms: D002460:Cell Line; D004268:DNA-Binding Proteins; D034441:Heterogeneous-Nuclear Ribonucleoproteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism*; D054884:Host-Pathogen Interactions; D006801:Humans; D047468:Immunoprecipitation; D018613:Microscopy, Confocal; D010750:Phosphoproteins / Q000378:metabolism; D011485:Protein Binding; D025941:Protein Interaction Mapping; D016601:RNA-Binding Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism*; D018116:Vesiculovirus / Q000502:physiology*; D015678:Viral Structural Proteins / Q000378:metabolism; D014779:Virus Replication
chemical_list: D004268:DNA-Binding Proteins; D034441:Heterogeneous-Nuclear Ribonucleoproteins; C081250:P protein, Vesicular stomatitis virus; C094604:PCBP1 protein, human; C501903:PCBP2 protein, human; D010750:Phosphoproteins; D016601:RNA-Binding Proteins; D015678:Viral Structural Proteins