Document Detail


In vivo and in vitro phosphorylation of rotavirus NSP5 correlates with its localization in viroplasms.
MedLine Citation:
PMID:  8985320     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
NSP5 (NS26), the product of rotavirus gene 11, is a phosphoprotein whose role in the virus replication cycle is unknown. To gain further insight into its function, we obtained monoclonal antibodies against the baculovirus-expressed protein. By immunoprecipitation and immunoblotting experiments, we showed that (i) NSP5 appears in many different phosphorylated forms in rotavirus-infected cells; (ii) immunoprecipitated NSP5 from rotavirus-infected cells can be phosphorylated in vitro by incubation with ATP; (iii) NSP5, produced either by transient transfection of rotavirus gene 11 or by infection by gene 11 recombinant vaccinia virus or baculovirus, can be phosphorylated in vivo and in vitro; (iv) NSP5 expressed in Escherichia coli is phosphorylated in vitro, and thus NSP5 is a potential protein kinase; and (v) NSP5 forms dimers and interacts with NSP2. The intracellular localization of NSP5 in the course of rotavirus infection and after transient expression in COS7 cells has also been investigated. In rotavirus-infected cells, NSP5 is localized in viroplasms, but it is widespread throughout the cytoplasm of transfected COS7 cells. NSP5 produced by transfected COS7 cells did not acquire the multiphosphorylated forms observed in rotavirus-infected COS7 cells. Thus, there is a tight correlation between the localization of NSP5 in the viroplasms and its protein kinase activity in vivo or in vitro. Our results suggest that cellular or viral cofactors are indispensable to fully phosphorylate NSP5 and to reach its intracellular localization.
Authors:
D Poncet; P Lindenbaum; R L'Haridon; J Cohen
Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Journal of virology     Volume:  71     ISSN:  0022-538X     ISO Abbreviation:  J. Virol.     Publication Date:  1997 Jan 
Date Detail:
Created Date:  1997-01-31     Completed Date:  1997-01-31     Revised Date:  2009-11-19    
Medline Journal Info:
Nlm Unique ID:  0113724     Medline TA:  J Virol     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  34-41     Citation Subset:  IM    
Affiliation:
Laboratoire de Virologie et Immunologie Moléculaires INRA, CRJJ, Jouy-en-Josas, France. poncet@biotec.jouy.inra.fr
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MeSH Terms
Descriptor/Qualifier:
Animals
COS Cells
Cell Line
Guinea Pigs
Mice
Mice, Inbred BALB C
Phosphorylation
Protein Kinases / metabolism*
RNA-Binding Proteins / metabolism
Rotavirus / metabolism*
Viral Nonstructural Proteins / metabolism
Viral Proteins / metabolism*
Chemical
Reg. No./Substance:
0/RNA-Binding Proteins; 0/Viral Nonstructural Proteins; 0/Viral Proteins; 138414-65-0/NS35 protein, rotavirus; 145715-40-8/gene 11 protein, Rotavirus B; EC 2.7.-/Protein Kinases
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