Identification of the nucleotide sequences of the oligonucleotides present in reovirions

Nichols, J. L.; Bellamy, A R; Joklik, Wolfgang K.

doi:10.1016/0042-6822(72)90507-7

Cited by 49 publications

(24 citation statements)

References 24 publications

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“…It is possible that the location of both 2 and 2 at the icosahedral vertices (7, 11) influences particle stability. Virions contain not only genomic dsRNA, a potential RIG-I and MDA5 ligand, but also short oligonucleotides with free 5= triphosphates (3,34,43), an additional potential RIG-I ligand, although shorter than other RIG-I agonists (27,28). Therefore, T3D particles may aberrantly disassemble more readily than T1L particles, resulting in commensurately greater PRR activation and induction of IFN.…”

Section: Discussionmentioning

confidence: 99%

A Single-Amino-Acid Polymorphism in Reovirus Protein μ2 Determines Repression of Interferon Signaling and Modulates Myocarditis

Irvin

Zurney

Ooms

et al. 2012

J Virol

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Section: Discussionmentioning

confidence: 99%

A Single-Amino-Acid Polymorphism in Reovirus Protein μ2 Determines Repression of Interferon Signaling and Modulates Myocarditis

Irvin

Zurney

Ooms

et al. 2012

J Virol

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“…A study of immature progeny virions isolated from reovirus-infected cells indicated that synthesis of these particle-associated RNA oligos occurs at a late step in virion morphogenesis (32). It was hypothesized that the RNA oligos, which comprise both abortive transcripts and poly(A), are products of the reovirus polymerase after outer capsid assembly (29,33,34). Although there is evidence that RNA oligos are not required for infection (35), their significance has not been investigated further.…”

Section: -7)mentioning

confidence: 99%

Transcriptional Activities of Reovirus RNA Polymerase in Recoated Cores

Farsetta¹,

Chandran²,

Nibert³

2000

Journal of Biological Chemistry

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The particle-associated reovirus polymerase synthesizes mRNA within only certain viral particle types. Reovirus cores, subviral particles lacking outer capsid proteins 1, 3, and 1, produce mRNA and abortive transcripts. Reovirus virions, which contain complete outer capsids, cannot produce mRNA and produce few abortive transcripts. Recoated cores are virion-like particles generated by the addition of recombinant outer capsid proteins to cores. We used recoated cores to analyze transcriptional regulation by reovirus outer capsid proteins. Partially recoated particles, containing less than virion amounts of 1 and 3, synthesized mRNA at levels inversely proportional to outer capsid protein levels. Fully recoated cores exhibited undetectable mRNA synthesis levels, as did virions. However, recoated cores produced high levels of abortive transcripts. Recoated core abortive transcripts remained particle-associated and appeared to inhibit further abortive transcript production. Proteolysis of recoated cores removing 1 and 3 released accumulated abortive transcripts and relieved inhibition of mRNA and abortive transcript synthesis. These results suggest transcriptional elongation, but not initiation, is blocked by virion-like amounts of 1 and 3. Particle-associated abortive transcripts may down-regulate transcriptional initiation. Minor outer capsid protein 1 had no demonstrable effect on transcriptional activities. Transcriptional regulation may ensure progeny virions do not compete with transcribing particles for ribonucleoside triphosphates.Both cellular and viral RNA polymerases synthesize abortive transcripts in addition to full-length mRNA (discussed in Refs. 1-7). This leads to the conceptual division of transcription into two basic stages: initiation and elongation. Initiation alone yields abortive transcripts, whereas initiation followed by elongation yields full-length mRNA. A body of work identifying physical differences between initiating and elongating transcription complexes supports this two-stage model. For example, the carboxyl-terminal domain of cellular RNA polymerase II is unphosphorylated during initiation but highly phosphorylated during elongation (8 -11). The phosphorylation state of P protein in the respiratory syncytial virus polymerase complex is important for the transition from initiation to elongation (12). Human immunodeficiency virus Tat protein is required for synthesis of full-length viral transcripts but not abortive transcripts (reviewed in Ref. 13). Certain mutations of bacteriophage T7 RNA polymerase render it capable of initiation but not elongation (14, 15). These findings suggest that RNA polymerases are regulated separately at initiation and elongation stages.Mammalian orthoreovirus (reovirus) has been used as a model to study transcription since its particle-associated RNAdependent RNA polymerase efficiently synthesizes large amounts of full-length mRNA (16 -20). Other particle-associated viral enzymes modify viral transcripts through the addition of the eukaryotic 5Ј-cap struct...

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“…Total RNA was obtained from the poly(A) polymerase reaction mixture by lysing the virions with 0.5 % SDS followed by two extractions with water-saturated phenol, and three additional extractions of the final aqueous phase with ether, to remove residual phenol (Nichols et al, 1972). Samples were then applied to 2.5 × 15 cm Sephadex G-100 columns, equilibrated with 1 mM-EDTA, 20 mM-tris pH 8 and eluted at 0 °C with the same buffer at a flow rate of 10 ml/h.…”

Section: Fraction Numbermentioning

confidence: 99%

“…The RNA isolated from reovirus (Fig. 2 b) eluted in two peaks, one in the excluded volume, corresponding to double-stranded genomic RNA, and a second one corresponding to the single-stranded oligonucleotides (Nichols et aL, 1972;Stoltzfus & Banerjee, 1972). The poly(A)-containing product of the in vitro reaction (Fig.…”

Section: Fraction Numbermentioning

confidence: 99%