Document Detail

Mapping of the RNA-binding domain of the cucumber mosaic virus movement protein.
MedLine Citation:
PMID:  9267013     Owner:  NLM     Status:  MEDLINE    
A series of in-frame deletion mutants was used to identify a domain within the 3a protein of cucumber mosaic virus (CMV) that is required for RNA-binding activity. Deletions in the 3a gene were generated by PCR and restriction digestion, and the resulting mutated 3a sequences were cloned either in pT7-7 or in pGEX-5X3 expression vectors. The mutated 3a proteins or fusions with glutathione S-transferase (GST) were expressed in E. coli, purified, and their nucleic acid-binding activities analysed by photochemical UV cross-linking assays using digoxigenin-UTP-labelled RNA probes. Comparative analyses of seven mutated 3a proteins obtained from inclusion bodies and eight GST fusion proteins revealed that there is an RNA-binding domain located between amino acids 174 and 233. This RNA-binding domain is able to bind single-stranded RNA out of the context of the complete 3a movement protein and is highly conserved within both subgroups of CMV.
C Vaquero; Y C Liao; J Nähring; R Fischer
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  The Journal of general virology     Volume:  78 ( Pt 8)     ISSN:  0022-1317     ISO Abbreviation:  J. Gen. Virol.     Publication Date:  1997 Aug 
Date Detail:
Created Date:  1997-09-08     Completed Date:  1997-09-08     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  0077340     Medline TA:  J Gen Virol     Country:  ENGLAND    
Other Details:
Languages:  eng     Pagination:  2095-9     Citation Subset:  IM    
Institut fur Biologie I (Molekulargenetik/Botanik), RWTH Aachen, Germany.
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MeSH Terms
Amino Acid Sequence
Binding Sites
Cucumovirus / metabolism*
Glutathione Transferase
Molecular Sequence Data
Mutagenesis, Site-Directed
Plant Viral Movement Proteins
Polymerase Chain Reaction
RNA, Viral / metabolism
RNA-Binding Proteins / chemistry*,  metabolism*
Recombinant Fusion Proteins / chemistry,  metabolism
Sequence Deletion
Ultraviolet Rays
Viral Proteins / chemistry*,  metabolism*
Reg. No./Substance:
0/Plant Viral Movement Proteins; 0/RNA, Viral; 0/RNA-Binding Proteins; 0/Recombinant Fusion Proteins; 0/Viral Proteins; EC Transferase

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