title: The respiratory syncytial virus polymerase has multiple RNA synthesis activities at the promoter.
abstract: Respiratory syncytial virus (RSV) is an RNA virus in the Family Paramyxoviridae. Here, the activities performed by the RSV polymerase when it encounters the viral antigenomic promoter were examined. RSV RNA synthesis was reconstituted in vitro using recombinant, isolated polymerase and an RNA oligonucleotide template representing nucleotides 1-25 of the trailer complement (TrC) promoter. The RSV polymerase was found to have two RNA synthesis activities, initiating RNA synthesis from the +3 site on the promoter, and adding a specific sequence of nucleotides to the 3' end of the TrC RNA using a back-priming mechanism. Examination of viral RNA isolated from RSV infected cells identified RNAs initiated at the +3 site on the TrC promoter, in addition to the expected +1 site, and showed that a significant proportion of antigenome RNAs contained specific nucleotide additions at the 3' end, demonstrating that the observations made in vitro reflected events that occur during RSV infection. Analysis of the impact of the 3' terminal extension on promoter activity indicated that it can inhibit RNA synthesis initiation. These findings indicate that RSV polymerase-promoter interactions are more complex than previously thought and suggest that there might be sophisticated mechanisms for regulating promoter activity during infection.
authors.affiliation: Department of Microbiology, Boston University School of Medicine, Boston, Massachusetts, USA.
authors.forename: Sarah L
authors.initials: SL
authors.lastname: Noton
authors.forename: Laure R
authors.initials: LR
authors.lastname: Deflubé
authors.forename: Chadene Z
authors.initials: CZ
authors.lastname: Tremaglio
authors.forename: Rachel
authors.initials: R
authors.lastname: Fearns
mesh_terms: D002460:Cell Line; D016679:Genome, Viral; D006801:Humans; D011401:Promoter Regions, Genetic; D018075:RNA, Complementary; D012367:RNA, Viral / Q000096:biosynthesis*; D012324:RNA-Dependent RNA Polymerase / Q000378:metabolism*; D018357:Respiratory Syncytial Virus Infections / Q000821:virology; D012136:Respiratory Syncytial Viruses / Q000201:enzymology* / Q000235:genetics* / Q000378:metabolism; D014158:Transcription, Genetic; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D018075:RNA, Complementary; D012367:RNA, Viral; D014764:Viral Proteins; D012324:RNA-Dependent RNA Polymerase