Promoter analysis of influenza virus RNA polymerase

J. D. Parvin, P. Palese, A. Honda, A. Ishihama, M. Krystal

Research output: Contribution to journalArticlepeer-review

145 Scopus citations

Abstract

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.

Original languageEnglish
Pages (from-to)5142-5152
Number of pages11
JournalJournal of Virology
Volume63
Issue number12
DOIs
StatePublished - 1989

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