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PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
9
pubmed:dateCreated
1995-9-14
pubmed:abstractText
Previously, a cDNA was constructed so that transcription by T7 RNA polymerase yielded a approximately 1-kb negative-sense analog of genomic RNA of human respiratory syncytial virus (RSV) containing the gene for chloramphenicol acetyltransferase (CAT) under the control of putative RSV transcription motifs and flanked by the RSV genomic termini. When transfected into RSV-infected cells, this minigenome was "rescued," as evidenced by high levels of CAT expression and the production of transmissible particles which propagated and expressed high levels of CAT expression during serial passage (P.L. Collins, M. A. Mink, and D. S. Stec, Proc. Natl. Acad. Sci. USA, 88:9663-9667, 1991). Here, this cDNA, together with a second one designed to yield an exact-copy positive-sense RSV-CAT RNA antigenome, were each modified to contain a self-cleaving hammerhead ribozyme for the generation of a nearly exact 3' end. Each cDNA was transfected into cells infected with a vaccinia virus recombinant expressing T7 RNA polymerase, together with plasmids encoding the RSV N, P, and L proteins, each under the control of a T7 promoter. When the plasmid-supplied template was the mini-antigenome, the minigenome was produced. When the plasmid-supplied template was the minigenome, the products were mini-antigenome, subgenomic polyadenylated mRNA and progeny minigenome. Identification of progeny minigenome made from the plasmid-supplied minigenome template indicates that the full RSV RNA replication cycle occurred. RNA synthesis required all three RSV proteins, N, P, and L, and was ablated completely by the substitution of Asn for Asp at position 989 in the L protein. Thus, the N, P, and L proteins were sufficient for the synthesis of correct minigenome and antigenome, but this was not the case for subgenomic mRNA, indicating that the requirements for RNA replication and transcription are not identical. Complementation with N, P, and L alone yielded an mRNA pattern containing a large fraction of molecules of incomplete, heterogeneous size. In contrast, complementation with RSV (supplying all of the RSV gene products) yielded a single discrete mRNA band. Superinfection with RSV of cells staging N/P/L-based RNA synthesis yielded the single discrete mRNA species. Some additional factor supplied by RSV superinfection appeared to be involved in transcription, the most obvious possibility being one or more additional RSV gene products.
pubmed:commentsCorrections
http://linkedlifedata.com/resource/pubmed/commentcorrection/7637014-1313041, http://linkedlifedata.com/resource/pubmed/commentcorrection/7637014-1318785, http://linkedlifedata.com/resource/pubmed/commentcorrection/7637014-1404620, http://linkedlifedata.com/resource/pubmed/commentcorrection/7637014-1621951, http://linkedlifedata.com/resource/pubmed/commentcorrection/7637014-1633429, http://linkedlifedata.com/resource/pubmed/commentcorrection/7637014-1648220, http://linkedlifedata.com/resource/pubmed/commentcorrection/7637014-1670500, http://linkedlifedata.com/resource/pubmed/commentcorrection/7637014-1840712, http://linkedlifedata.com/resource/pubmed/commentcorrection/7637014-1946383, http://linkedlifedata.com/resource/pubmed/commentcorrection/7637014-2053282, http://linkedlifedata.com/resource/pubmed/commentcorrection/7637014-2107573, http://linkedlifedata.com/resource/pubmed/commentcorrection/7637014-2161049, http://linkedlifedata.com/resource/pubmed/commentcorrection/7637014-2497259, http://linkedlifedata.com/resource/pubmed/commentcorrection/7637014-2548200, http://linkedlifedata.com/resource/pubmed/commentcorrection/7637014-2585601, http://linkedlifedata.com/resource/pubmed/commentcorrection/7637014-2598262, http://linkedlifedata.com/resource/pubmed/commentcorrection/7637014-3095828, http://linkedlifedata.com/resource/pubmed/commentcorrection/7637014-3476956, http://linkedlifedata.com/resource/pubmed/commentcorrection/7637014-3757031, http://linkedlifedata.com/resource/pubmed/commentcorrection/7637014-3839952, http://linkedlifedata.com/resource/pubmed/commentcorrection/7637014-6548527, http://linkedlifedata.com/resource/pubmed/commentcorrection/7637014-6960240, http://linkedlifedata.com/resource/pubmed/commentcorrection/7637014-7884888, http://linkedlifedata.com/resource/pubmed/commentcorrection/7637014-7925265, http://linkedlifedata.com/resource/pubmed/commentcorrection/7637014-7975205, http://linkedlifedata.com/resource/pubmed/commentcorrection/7637014-7975856, http://linkedlifedata.com/resource/pubmed/commentcorrection/7637014-8078927, http://linkedlifedata.com/resource/pubmed/commentcorrection/7637014-8091846, http://linkedlifedata.com/resource/pubmed/commentcorrection/7637014-8257115, http://linkedlifedata.com/resource/pubmed/commentcorrection/7637014-8277263, http://linkedlifedata.com/resource/pubmed/commentcorrection/7637014-8289375, http://linkedlifedata.com/resource/pubmed/commentcorrection/7637014-8317100, http://linkedlifedata.com/resource/pubmed/commentcorrection/7637014-8386276, http://linkedlifedata.com/resource/pubmed/commentcorrection/7637014-8389932, http://linkedlifedata.com/resource/pubmed/commentcorrection/7637014-8392616, http://linkedlifedata.com/resource/pubmed/commentcorrection/7637014-8395122, http://linkedlifedata.com/resource/pubmed/commentcorrection/7637014-8460489
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
http://linkedlifedata.com/resource/pubmed/chemical/Chloramphenicol O-Acetyltransferase, http://linkedlifedata.com/resource/pubmed/chemical/DNA-Directed RNA Polymerases, http://linkedlifedata.com/resource/pubmed/chemical/HN Protein, http://linkedlifedata.com/resource/pubmed/chemical/RNA, Catalytic, http://linkedlifedata.com/resource/pubmed/chemical/RNA, Messenger, http://linkedlifedata.com/resource/pubmed/chemical/RNA, Viral, http://linkedlifedata.com/resource/pubmed/chemical/Recombinant Proteins, http://linkedlifedata.com/resource/pubmed/chemical/Viral Core Proteins, http://linkedlifedata.com/resource/pubmed/chemical/Viral Envelope Proteins, http://linkedlifedata.com/resource/pubmed/chemical/Viral Proteins, http://linkedlifedata.com/resource/pubmed/chemical/attachment protein G, http://linkedlifedata.com/resource/pubmed/chemical/bacteriophage T7 RNA polymerase
pubmed:status
MEDLINE
pubmed:month
Sep
pubmed:issn
0022-538X
pubmed:author
pubmed:issnType
Print
pubmed:volume
69
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
5677-86
pubmed:dateRevised
2009-11-18
pubmed:meshHeading
pubmed-meshheading:7637014-Humans, pubmed-meshheading:7637014-Vaccinia virus, pubmed-meshheading:7637014-Base Sequence, pubmed-meshheading:7637014-RNA, Viral, pubmed-meshheading:7637014-Viral Proteins, pubmed-meshheading:7637014-RNA, Messenger, pubmed-meshheading:7637014-Respiratory Syncytial Viruses, pubmed-meshheading:7637014-Cell Line, pubmed-meshheading:7637014-Molecular Sequence Data, pubmed-meshheading:7637014-DNA-Directed RNA Polymerases, pubmed-meshheading:7637014-Transcription, Genetic, pubmed-meshheading:7637014-RNA, Catalytic, pubmed-meshheading:7637014-Genome, Viral, pubmed-meshheading:7637014-Bacteriophage T7, pubmed-meshheading:7637014-Capsid, pubmed-meshheading:7637014-Promoter Regions, Genetic, pubmed-meshheading:7637014-Terminator Regions, Genetic, pubmed-meshheading:7637014-Transfection
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