Identification of the RNA region transferred from a representative primer, beta-globin mRNA, to influenza mRNA during in vitro transcription

Abstract

Capped eukaryotic mRNAs strongly stimulate influenza viral RNA transcription in vitro and donate their cap and also additional nucleotides to the viral transcripts (1). To identify which bases of a given primer mRNA are transferred, we synthesized influenza viral mRNA using a primer rabbit globin mRNA (enriched in beta-globin mRNA) which had been labeled in vitro to high specific activity with 125I. We show that during transcription the same 125I-labeled oligonucleotides were transferred to the 5' termini of each of the eight viral mRNA segments. The predominant sequence, representing 75 percent of the transferred oligonucleotides, was identical to the first 13 nucleotides at the 5' end of beta-globin mRNA (m7G5'ppp5'm6AmC(m)ACUUGCUUUUG). Because only the C-residues are labeled with 125I, these results indicate that either the first 12, 13 or 14 5' terminal bases of beta-globin mRNA were transferred to the viral mRNAs. 125I-labeled oligonucleotides recovered from the viral mRNA in minor yields indicated that shorter 5' terminal pieces of beta-globin mRNA were sometimes transferred and that G was probably the first base inserted by transcription.