| Transcriptional repression and RNA silencing act synergistically to demonstrate the function of the eleventh component of the vaccinia virus entry-fusion complex. | |
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MedLine Citation:
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PMID: 22013036 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Poxviruses have an elaborate system for infecting cells comprising several proteins for attachment and a larger number dedicated to membrane fusion and entry. Thus far, 11 proteins have been identified as components of the vaccinia virus (VACV) entry-fusion complex (EFC), and 10 of these proteins have been shown to be required for entry. J5, the remaining functionally uncharacterized component of the complex, is conserved in all poxviruses, has a predicted C-terminal transmembrane domain, and is an N-terminally truncated paralog of two other EFC proteins. To determine the role of J5, we constructed a mutant that inducibly regulates J5 transcription. Although the virus yield was reduced only about 80% without inducer, the inability to isolate a J5 deletion mutant suggested an essential function. To enhance stringency, we employed RNA silencing alone and together with transcriptional repression of the inducible mutant. The yield of infectious virus was reduced 4- to 5-fold by repression, 2-fold by silencing, and 60-fold by the combination of the two. Virus particles made under the latter conditions appeared to contain a full complement of proteins excluding J5 but had very low infectivity. Further studies indicated that after binding to cells, J5-deficient virions had a defect in core entry and an inability to induce syncytium formation. In addition, we confirmed that J5 is associated with the EFC by affinity purification. These data indicate that J5 is a functional component of the EFC and highlights the advantage of combining transcriptional repression and RNA silencing for stringent reduction of gene expression. |
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Authors:
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Cindy L Wolfe; Suany Ojeda; Bernard Moss |
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Publication Detail:
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Type: Journal Article; Research Support, N.I.H., Intramural Date: 2011-10-19 |
Journal Detail:
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Title: Journal of virology Volume: 86 ISSN: 1098-5514 ISO Abbreviation: J. Virol. Publication Date: 2012 Jan |
Date Detail:
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Created Date: 2011-12-14 Completed Date: 2012-02-14 Revised Date: 2012-07-02 |
Medline Journal Info:
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Nlm Unique ID: 0113724 Medline TA: J Virol Country: United States |
Other Details:
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Languages: eng Pagination: 293-301 Citation Subset: IM |
Affiliation:
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Laboratory of Viral Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland 20892-3210, USA. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Amino Acid Sequence Cell Line Down-Regulation* Gene Expression Regulation, Viral Humans Molecular Sequence Data RNA Interference* Sequence Alignment Transcription, Genetic* Vaccinia / virology* Vaccinia virus / chemistry, genetics*, metabolism Viral Proteins / chemistry, genetics*, metabolism* |
| Chemical | |
Reg. No./Substance:
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0/Viral Proteins |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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