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. 1979 Dec;32(3):895–904. doi: 10.1128/jvi.32.3.895-904.1979

Cap and Internal Nucleotides of Reovirus mRNA Primers Are Incorporated into Influenza Viral Complementary RNA During Transcription In Vitro

Michele Bouloy 1, Maureen A Morgan 2, Aaron J Shatkin 2, Robert M Krug 1
PMCID: PMC525938  PMID: 513205

Abstract

Reovirus mRNA's containing a 5′-terminal methylated cap structure (m7GpppGm) were shown to be effective primers for influenza viral RNA transcription in vitro catalyzed by the influenza virion transcriptase. Priming activity required the presence of methyl groups in the cap since reovirus mRNA's with 5′-terminal GpppG were inactive as primers. Both the cap and internal nucleotides were physically transferred from radiolabeled reovirus mRNA to influenza viral complementary RNA (cRNA) during transcription in vitro. By using reovirus mRNA's with methyl-3H-labeled caps as primers, we showed that the influenza viral cRNA synthesized in the presence of unlabeled nucleoside triphosphates contained [methyl-3H]m7GpppGm, identical to that found in the reovirus mRNA primer. To demonstrate transfer of internal residues, reovirus mRNA's synthesized in the presence of all four α-32P-labeled ribonucleoside triphosphates were used as primers. The resulting influenza viral cRNA was 32P-labeled. Diethyl-aminoethyl-Sephadex chromatography of the RNase T2 digest of this cRNA demonstrated 32P radiolabel in both internal residues (charge −2) and the cap (charge −4.6). Approximately 25 internal nucleotides along with the cap of reovirus mRNA were transferred to each chain of influenza viral cRNA. Gel electrophoretic analysis indicated that the segments of influenza viral cRNA primed by reovirus mRNA were approximately the same size as those primed by a different mRNA, globin mRNA, strongly suggesting that the influenza virion transcriptase complex transfers approximately the same number of nucleotides plus the cap from different mRNA primers to the 5′ end of influenza viral RNA transcripts.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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