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| Predicate | Object |
|---|---|
| rdf:type | |
| lifeskim:mentions | |
| pubmed:issue |
7
|
| pubmed:dateCreated |
1998-7-1
|
| pubmed:abstractText |
We developed a system to identify the viral proteins required for the packaging and passage of human respiratory syncytial virus (RSV) by reconstructing these events with cDNA-encoded components. Plasmids encoding individual RSV proteins, each under the control of a T7 promoter, were cotransfected in various combinations together with a plasmid containing a minigenome into cells infected with a vaccinia virus recombinant expressing T7 RNA polymerase. Supernatants from these cells were passaged onto fresh cells which were then superinfected with RSV. Functional reconstitution of RSV-specific packaging and passage was detected by expression of the reporter gene carried on the minigenome. As expected, the four nucleocapsid proteins N, P, L, and M2-1 failed to direct packaging and passage of the minigenome. Passage was achieved by further addition of plasmids expressing three membrane-associated proteins, M, G, and F; inclusion of the fourth envelope- associated protein, SH, did not alter passage efficiency. Passage was reduced 10- to 20-fold by omission of G and was abrogated by omission of either M or F. Coexpression of the nonstructural NS1 or NS2 protein had little effect on packaging and passage except through indirect effects on RNA synthesis in the initial transfection. The M2-1 transcription elongation factor was not required for the generation of passage-competent particles. However, addition of increasing quantities of M2-1 to the transfection mediated a dose-dependent inhibition of passage which was alleviated by coexpression of the putative negative regulatory factor M2-2. Omission of the L plasmid reduced passage 10- to 20-fold, most likely due to reduced availability of encapsidated minigenomes for packaging. However, the residual level of passage indicated that neither L protein nor the process of RSV-specific RNA synthesis is required for the production and passage of particles. Omission of N or P from the transfection abrogated passage. Thus, the minimum RSV protein requirements for packaging and passaging a minigenome are N, P, M, and F, although the efficiency is greatly increased by addition of L and G.
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| pubmed:commentsCorrections |
http://linkedlifedata.com/resource/pubmed/commentcorrection/9621029-1318785,
http://linkedlifedata.com/resource/pubmed/commentcorrection/9621029-1847519,
http://linkedlifedata.com/resource/pubmed/commentcorrection/9621029-2011192,
http://linkedlifedata.com/resource/pubmed/commentcorrection/9621029-2547986,
http://linkedlifedata.com/resource/pubmed/commentcorrection/9621029-3532115,
http://linkedlifedata.com/resource/pubmed/commentcorrection/9621029-7637014,
http://linkedlifedata.com/resource/pubmed/commentcorrection/9621029-7707520,
http://linkedlifedata.com/resource/pubmed/commentcorrection/9621029-7793072,
http://linkedlifedata.com/resource/pubmed/commentcorrection/9621029-8093220,
http://linkedlifedata.com/resource/pubmed/commentcorrection/9621029-8178462,
http://linkedlifedata.com/resource/pubmed/commentcorrection/9621029-8289375,
http://linkedlifedata.com/resource/pubmed/commentcorrection/9621029-8552680,
http://linkedlifedata.com/resource/pubmed/commentcorrection/9621029-8601317,
http://linkedlifedata.com/resource/pubmed/commentcorrection/9621029-8764008,
http://linkedlifedata.com/resource/pubmed/commentcorrection/9621029-8876140,
http://linkedlifedata.com/resource/pubmed/commentcorrection/9621029-8876141,
http://linkedlifedata.com/resource/pubmed/commentcorrection/9621029-9266983,
http://linkedlifedata.com/resource/pubmed/commentcorrection/9621029-9298896,
http://linkedlifedata.com/resource/pubmed/commentcorrection/9621029-9298897,
http://linkedlifedata.com/resource/pubmed/commentcorrection/9621029-9299631,
http://linkedlifedata.com/resource/pubmed/commentcorrection/9621029-9371553,
http://linkedlifedata.com/resource/pubmed/commentcorrection/9621029-9391135,
http://linkedlifedata.com/resource/pubmed/commentcorrection/9621029-9420254,
http://linkedlifedata.com/resource/pubmed/commentcorrection/9621029-9445048,
http://linkedlifedata.com/resource/pubmed/commentcorrection/9621029-9499071
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| pubmed:language |
eng
|
| pubmed:journal | |
| pubmed:citationSubset |
IM
|
| pubmed:chemical | |
| pubmed:status |
MEDLINE
|
| pubmed:month |
Jul
|
| pubmed:issn |
0022-538X
|
| pubmed:author | |
| pubmed:issnType |
Print
|
| pubmed:volume |
72
|
| pubmed:owner |
NLM
|
| pubmed:authorsComplete |
Y
|
| pubmed:pagination |
5707-16
|
| pubmed:dateRevised |
2009-11-18
|
| pubmed:meshHeading |
pubmed-meshheading:9621029-Genome, Viral,
pubmed-meshheading:9621029-Humans,
pubmed-meshheading:9621029-RNA, Viral,
pubmed-meshheading:9621029-Respiratory Syncytial Virus, Human,
pubmed-meshheading:9621029-Viral Nonstructural Proteins,
pubmed-meshheading:9621029-Viral Proteins,
pubmed-meshheading:9621029-Virion,
pubmed-meshheading:9621029-Virus Assembly
|
| pubmed:year |
1998
|
| pubmed:articleTitle |
Identification of the respiratory syncytial virus proteins required for formation and passage of helper-dependent infectious particles.
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| pubmed:affiliation |
Laboratory of Infectious Diseases, National Institute of Allergy and Infectious Diseases, Bethesda, Maryland 20892-0720, USA.
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| pubmed:publicationType |
Journal Article
|