Isolation and Properties of the Replicase of Encephalomyocarditis Virus

Traub, A.; Diskin, B.; Rosenberg, H.; Kalmar, E.

doi:10.1128/jvi.18.2.375-382.1976

Cited by 45 publications

(21 citation statements)

References 31 publications

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“…These experiments provide strong evidence that EMC RNA codes for an active proteolytic enzyme, with an essential -SH group, which can cleave pre-A and presumably C. This enzyme is not contained in pre-A, as this is stable on its own. Since D and E are found in purified preparations of the viral replicase [28], and are themselves generated by secondary cleavage, I and F would seem to be likely candidates for the protease. I does seem to be a primary product [26], since it is not produced by cleavage of pre-A (Fig.…”

Section: Resultsmentioning

confidence: 99%

Translation of Encephalomyocarditis Virus RNA in vitro Yields an Active Proteolytic Processing Enzyme

Pelham

1978

European Journal of Biochemistry

155

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In contrast to other cell‐free translation systems, the mRNA‐dependent reticulocyte lysate can translate encephalomyocarditis virus RNA efficiently and completely when supplemented with heterologous tRNA. Cleavage of the nascent polypeptide chain occurs, and one of the translation products appears to be a specific proteolytic enzyme which correctly processes the primary products. The identity of the proteins made in vitro was verified by comparison with infected cell proteins on dodecylsulphate/polyacrylamide gels, and by mapping their coding sequences on the viral genome.

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Section: Resultsmentioning

confidence: 99%

Translation of Encephalomyocarditis Virus RNA in vitro Yields an Active Proteolytic Processing Enzyme

Pelham

1978

European Journal of Biochemistry

155

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“…The present results show that a 56kilodalton protein synthesized in FMDV-infected cells forms part of a 70S complex which functions as a poly(U) polymerase. Enzyme complexes isolated from other picornaviruses have been reported to contain a virus-specific polypeptide having a molecular weight of about 56,000 (12,14,22).…”

Section: Vol 40 1981mentioning

confidence: 99%

Characterization of a 70S polyuridylic acid polymerase isolated from foot-and-mouth disease virus-infected cells

Polatnick¹,

Wool²

1981

J Virol

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A polyuridylic acid polymerase complex isolated from foot-and-mouth disease virus-infected cells sedimented at 70S in a sucrose gradient and appeared in the exclusion volume of an agarose column whose molecular weight cutoff was 5 x 10(6). Phenol extraction of the complex yielded a heterogeneous band of virus-specific RNA and an apparently host cell-derived 4.5 to 5S RNA, both of which are essentially single stranded. Neither RNA served as a template in the cell-free enzyme reaction. Polyacrylamide gel analysis revealed five polypeptides with molecular weights of 50,000, 56,000, 60,000, 70,000, and 74,000 and with molar ratios of 1:2:2:1:1, respectively. Autoradiography showed P56 to be the only major virus-induced polypeptide; the other proteins are apparently of host cell origin. Electron microscopic examination suggested a cartwheel shape for the polymerase complex which was seen to dissociate as polyadenylic acid was added. Antibody previously shown to inhibit enzyme activity aggregated the 70S units.

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“…(3) To eliminate all potential translation initiation codons (65,66) upstream from and in-frame with the "INS"/laminin sequence in the edited sense strand RNA; (4) To introduce within the TCE of the projected antisense strand an AUG codon in the 5' to 3' orientation, in the optimal translation initiation context, and inframe with the "INS"/ laminin coding sequence in the projected chimeric RNA end product of amplification; (5) To minimize TCE/ICE mismatches so as to assure that the cleavage of a chimeric intermediate does not occur downstream from the "AUG" and to maximize the length of the 5'UTR in the projected chimeric end product; (6) To ascertain that the "AUG" of the antisense RNA is followed by a codon encoding Val so as to confer to the resulting protein the maximum half-life, in accordance with the N-end rule pathway of protein degradation (67), if the terminal Met is removed by the N-terminal methionyl aminopeptidase (68); (7) To minimally interfere with the 5'terminus of the sense strand transcript so as to preserve the position of the TSS. Stages C and D of the middle panel of Figure 12 show the folding of the antisense RNA (stage C) and its extension into chimeric intermediate followed by strand separation and cleavage (red arrow, stage D).…”

Section: Regulatory Aspects Of Mammalian Mrna Amplification Processmentioning

confidence: 99%

Protein-Encoding RNA-to-RNA Information Transfer in Mammalian Cells: Principles of RNA-Dependent mRNA Amplification

Volloch¹

2019

Preprint

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The transfer of protein-encoding genetic information from DNA to RNA to protein, a process formalized as the "Central Dogma of Molecular Biology", has undergone a significant evolution since its inception. It was amended to account for the information flow from RNA to DNA, the reverse transcription, and for the information transfer from RNA to RNA, the RNA-dependent RNA synthesis. These processes, both potentially leading to protein production, were initially described only in viral systems, and although RNAdependent RNA polymerase activity was shown to be present, and RNA-dependent RNA synthesis found to occur in mammalian cells, its function was presumed to be restricted to regulatory. However, recent results, obtained in several systems, strongly indicate the occurrence of protein-encoding RNA to RNA information transfer in mammalian cells. It can result in the rapid production of the extraordinary quantities of specific proteins as was seen in cases of terminal cellular differentiation and during cellular deposition of extracellular matrix molecules. A malfunction of this process may be involved in pathologies associated either with the deficiency of a protein normally produced by this mechanism or with the abnormal abundance of a protein or of its C-terminal fragment. It seems to be responsible for some types of familial thalassemia and may underlie the overproduction of beta amyloid in sporadic Alzheimer's disease. The increased understanding of components and mechanisms involved in RNA-dependent mammalian mRNA amplification could open up new approaches not only for therapeutic interference in multiple pathologies but also for novel and powerful forms of bioengineering. The aim of the present article is to systematize the current knowledge and understanding of this pathway. The outlined framework introduces unexpected features of the mRNA amplification such as its second Tier, a physiologically occurring intracellular PCR, iPCR, a "Two-Tier Paradox" and RNA "Dark Matter". It also describes novel experimental bioengineering designs and may serve as a basis for new directions of investigations into the mechanisms underlying the mammalian mRNA amplification processes.

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Isolation and Properties of the Replicase of Encephalomyocarditis Virus

Cited by 45 publications

References 31 publications

Translation of Encephalomyocarditis Virus RNA in vitro Yields an Active Proteolytic Processing Enzyme

Translation of Encephalomyocarditis Virus RNA in vitro Yields an Active Proteolytic Processing Enzyme

Characterization of a 70S polyuridylic acid polymerase isolated from foot-and-mouth disease virus-infected cells

Protein-Encoding RNA-to-RNA Information Transfer in Mammalian Cells: Principles of RNA-Dependent mRNA Amplification

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