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Nucleic Acids Research, 1980, Vol. 8, No. 5 925-942
© 1980

Articles

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

Hugh D. Robertson*, Elizabeth Dickson*, Stephen J. Plotch{dagger} and Robert M. Krug{dagger}

*The Rockefeller University New York, NY 10021 {dagger}Memorial Sloan-Kettering Cancer Center New York, NY 10021, USA

Received January 28, 1980. 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 ß-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 ß-globin mRNA (m7G5'ppp5' m6AmC(m)ACUUGCUUUUG). Because only the C-re-sidues are labeled with 125I, these results indicate that either the first 12, 13 or 14 5' terminal bases of ß-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 ß-globin mRNA were sometimes transferred and that G was probably the first base inserted by transcription.


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