title: Characterization of grass carp reovirus minor core protein VP4.
abstract: BACKGROUND
  Grass Carp Reovirus (GCRV), a tentative member in the genus Aquareovirus of family Reoviridae, contains eleven segmented (double-stranded RNA) dsRNA genome which encodes 12 proteins. A low-copy core component protein VP4, encoded by the viral genome segment 5(S5), has been suggested to play a key role in viral genome transcription and replication.
  
  
  RESULTS
  To understand the role of minor core protein VP4 played in molecular pathogenesis during GCRV infection, the recombinant GCRV VP4 gene was constructed and expressed in both prokaryotic and mammalian cells in this investigation. The recombinant His-tag fusion VP4 products expressed in E.coli were identified by Western blotting utilizing His-tag specific monoclonal and GCRV polyclonal antibodies. In addition, the expression of VP4 in GCRV infected cells, appeared in granules structure concentrated mainly in the cytoplasm, can be detected by Immunofluorescence (IF) using prepared anti-VP4 polyclonal antibody. Meanwhile, VP4 protein in GCRV core and infected cell lysate was identified by Immunoblotting (IB) assay. Of particular note, the VP4 protein was exhibited a diffuse distribution in the cytoplasm and nucleus in transfected cells, suggesting that VP4 protein may play a partial role in the nucleus by regulating cell cycle besides its predicted cytoplasmic function in GCRV infection.
  
  
  CONCLUSIONS
  Our results indicate the VP4 is a core component in GCRV. The cellular localization of VP4 is correlated with its predicted function. The data provide a foundation for further studies aimed at understanding the role of VP4 in viroplasmic inclusion bodies (VIB) formation during GCRV replication and assembly.
authors.affiliation: State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, 430071, China.
authors.forename: Liming
authors.initials: L
authors.lastname: Yan
authors.forename: Hong
authors.initials: H
authors.lastname: Guo
authors.forename: Xiaoyun
authors.initials: X
authors.lastname: Sun
authors.forename: Ling
authors.initials: L
authors.lastname: Shao
authors.forename: Qin
authors.initials: Q
authors.lastname: Fang
mesh_terms: D000818:Animals; D002460:Cell Line; D002467:Cell Nucleus / Q000737:chemistry; D003593:Cytoplasm / Q000737:chemistry; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D015151:Immunoblotting; D008856:Microscopy, Fluorescence; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D012087:Reoviridae / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D014758:Viral Core Proteins / Q000235:genetics / Q000378:metabolism*; D014779:Virus Replication
chemical_list: D011994:Recombinant Proteins; D014758:Viral Core Proteins