Polyadenylation and reverse transcription of influenza viral RNA

• Published in: Nucleic acids research Vol. 6; no. 4; pp. 1221 - 1239
• Main Authors: Emtage, J.S., Catlin, G.H., Carey, N.H.
• Format: Journal Article
• Language: English
• Published: England Oxford University Press 01-04-1979
• Subjects: DNA, Viral - biosynthesis; Escherichia coli; Escherichia coli - enzymology; Influenza A virus; Kinetics; Molecular Weight; Nucleic Acid Hybridization; Nucleotidyltransferases - metabolism; Orthomyxoviridae; Poly A - biosynthesis; Polynucleotide Adenylyltransferase - metabolism; RNA, Viral - metabolism; RNA-Directed DNA Polymerase - metabolism
• ISSN: 0305-1048; 1362-4962
• DOI: 10.1093/nar/6.4.1221

The polyadenylation of Fowl Plague Viral RNA and of Influenza A/Victoria Viral RNA using E.coli poly(A) polymerase and the subsequent reverse transcription of the polyadenylated species is reported. We have shown that all 8 genome fragments are adenylated and that an average of 25–30 adenylic acid residues per molecule is sufficient for maximal transcription with reverse transcriptase. The cDNA product is 95Z sensitive to S1-nuclease and hybridisation analysis against viral RNA reveals it to be a faithful copy of the RNA. Amongst the transcription products are long, discrete copies of genes 1–8, the lengths of which are comparable with those of the vRNA determined by electrophoresis on formamide acrylamide gels. These single-stranded cDNAs have been further transcribed to form double-stranded products with hair-pin structures at one end. Analysis of this material on native acrylamide gels revealed some DNA bands corresponding to the predicted sizes for genes 4–8.