Polyadenylation and reverse transcription of influenza viral RNA.
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 95% sensitive to Sl-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.
D004279:DNA, Viral / Q000096:biosynthesis; D004926:Escherichia coli / Q000201:enzymology*; D009980:Influenza A virus; D007700:Kinetics; D008970:Molecular Weight; D009693:Nucleic Acid Hybridization; D009713:Nucleotidyltransferases / Q000378:metabolism*; D009975:Orthomyxoviridae; D011061:Poly A / Q000096:biosynthesis*; D011062:Polynucleotide Adenylyltransferase / Q000378:metabolism*; D012367:RNA, Viral / Q000378:metabolism*; D012194:RNA-Directed DNA Polymerase / Q000378:metabolism*
D004279:DNA, Viral; D012367:RNA, Viral; D011061:Poly A; D009713:Nucleotidyltransferases; D011062:Polynucleotide Adenylyltransferase; D012194:RNA-Directed DNA Polymerase