| Mapping of the RNA-binding domain of the cucumber mosaic virus movement protein. | |
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MedLine Citation:
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PMID: 9267013 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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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. |
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Authors:
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C Vaquero; Y C Liao; J Nähring; R Fischer |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't |
Journal Detail:
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Title: The Journal of general virology Volume: 78 ( Pt 8) ISSN: 0022-1317 ISO Abbreviation: J. Gen. Virol. Publication Date: 1997 Aug |
Date Detail:
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Created Date: 1997-09-08 Completed Date: 1997-09-08 Revised Date: 2006-11-15 |
Medline Journal Info:
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Nlm Unique ID: 0077340 Medline TA: J Gen Virol Country: ENGLAND |
Other Details:
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Languages: eng Pagination: 2095-9 Citation Subset: IM |
Affiliation:
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Institut fur Biologie I (Molekulargenetik/Botanik), RWTH Aachen, Germany. |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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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* |
| Chemical | |
Reg. No./Substance:
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0/Plant Viral Movement Proteins; 0/RNA, Viral; 0/RNA-Binding Proteins; 0/Recombinant Fusion Proteins; 0/Viral Proteins; EC 2.5.1.18/Glutathione Transferase |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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