Promoter analysis of influenza virus RNA polymerase.
Influenza virus polymerase, which was prepared depleted of viral RNA, was used to copy small RNA templates prepared from plasmid-encoded sequences. Template constructions containing only the 3' end of genomic RNA were shown to be efficiently copied, indicating that the promoter lay solely within the 15-nucleotide 3' terminus. Sequences not specific for the influenza virus termini were not copied, and, surprisingly, RNAs containing termini identical to those from plus-sense cRNA were copied at low levels. The specificity for recognition of the virus sense promoter was further defined by site-specific mutagenesis. It was also found that increased levels of viral protein were required in order to catalyze both the cap endonuclease-primed and primer-free RNA synthesis from these model templates, as well as from genomic-length RNAs. This finding indicates that the reconstituted system has catalytic properties very similar to those of native viral ribonucleoprotein complexes.
Parvin
J D
JD
Department of Microbiology, Mount Sinai School of Medicine, New York, New York 10029.
Palese
P
P
Honda
A
A
Ishihama
A
A
Krystal
M
M
D001483:Base Sequence; D012321:DNA-Directed RNA Polymerases / Q000302:isolation & purification / Q000378:metabolism*; D005814:Genes, Viral; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D009975:Orthomyxoviridae / Q000235:genetics*; D010957:Plasmids; D011401:Promoter Regions, Genetic; D012367:RNA, Viral / Q000096:biosynthesis / Q000235:genetics*; D013698:Templates, Genetic; D014158:Transcription, Genetic
D015345:Oligonucleotide Probes; D012367:RNA, Viral; D012321:DNA-Directed RNA Polymerases