Absence of Detectable Capping and Methylating Enzymes in Influenza Virions

Plotch, Stephen J.; Tomasz, J.; Krug, Robert M.

doi:10.1128/jvi.28.1.75-83.1978

Cited by 34 publications

(13 citation statements)

References 47 publications

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“…Synthesis ofinfluenza viral eRNA in vitro. The synthesis of cRNA in vitro by detergent-treated, purified influenza virus was carried out as described previously (15,17). The primer and labeled ribonucleoside triphosphate added to each reaction mixture are indicated in each table and figure.…”

Section: Methodsmentioning

confidence: 99%

“…Enzymatic digestion and analytical methods. The conditions for digestion with P1 nuclease, BAP, nucleotide pyrophosphatase, and RNase T2 as well as analyses by high-voltage paper electrophoresis in pyridine acetate buffer (pH 3.5), paper chromatography in isobutyric solvents, and column chromatography on diethylaminoethyl (DEAE)-cellulose have been described previously (4,5,17).…”

Section: Methodsmentioning

confidence: 99%

“…The previous estimation that 10 to 15 nucleotides were transferred from a different primer, globin mRNA, to influenza viral cRNA was based on electrophoretic mobilities, specifically on the difference in mobility between the globin mRNA-primed and ApG-primed cRNA segments (14). The latter cRNA segments initiate exactly at the 3' end of the vRNA templates (17,20). To compare the number of nucleotides transferred to influenza viral cRNA from globin mRNA and reovirus mRNA, we compared the electrophoretic mobilities of the cRNA's primed by these mRNA's and ApG (Fig.…”

Section: Igimi* T?mentioning

confidence: 99%

“…Gel electrophoretic analysis of the globin mRNA-primed cRNA segments indicated that they were 10 to 15 nucleotides longer at their 5' end than the cRNA segments synthesized with ApG as primer. Since ApG has been shown to initiate transcription exactly at the 3' end of the virion RNA (vRNA) segments (17,20), it seemed likely that the 10 to 15 additional nucleotides were also physically transferred from the globin mRNA to the 5' end of viral cRNA during transcription (14).…”

mentioning

confidence: 99%

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

Bouloy

Morgan

Shatkin

et al. 1979

J Virol

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Reovirus mRNA's containing a 5′-terminal methylated cap structure (m 7 GpppG m ) 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 - 3 H-labeled caps as primers, we showed that the influenza viral cRNA synthesized in the presence of unlabeled nucleoside triphosphates contained [ methyl - 3 H]m 7 GpppG m , 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- 32 P-labeled ribonucleoside triphosphates were used as primers. The resulting influenza viral cRNA was 32 P-labeled. Diethyl-aminoethyl-Sephadex chromatography of the RNase T2 digest of this cRNA demonstrated 32 P 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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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Igimi* T?mentioning

confidence: 99%

mentioning

confidence: 99%

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

Bouloy

Morgan

Shatkin

et al. 1979

J Virol

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“…RIG-I, at least in murine bone marrow-derived dendritic cells (BM-DCs) and human HEK293 cells, has also been reported to detect IAV ssRNA that carries phosphorylated 59-ends (56). Unlike mammalian mRNA, the IAV viral genome comprises uncapped RNA segments with phosphate groups at their 59-termini (57,58). The ability of RIG-I to recognize these 59-phosphate groups on ssRNA and the dsRNA domains makes it a superior sensor.…”

Section: Rlrsmentioning

confidence: 99%