Biochemical analysis of the multifunctional vaccinia mRNA capping enzyme encoded by a temperature sensitive virus mutant

Abstract Prior biochemical analysis of the heterodimeric vaccinia virus mRNA capping enzyme suggests roles not only in mRNA capping but also in early viral gene transcription termination and intermediate viral gene transcription initiation. Prior phenotypic characterization of Dts36, a temperature s...

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Bibliographic Details
Published in:Virology (New York, N.Y.) N.Y.), 2016-01, Vol.487, p.27-40
Main Authors: Tate, Jessica, Boldt, Rachel L, McFadden, Baron D, D’Costa, Susan M, Lewandowski, Nicholas M, Shatzer, Amber N, Gollnick, Paul, Condit, Richard C
Format: Article
Language:English
Subjects:
Animals
Capping enzyme
Cell Line
Cercopithecus aethiops
HeLa Cells
Humans
Infectious Disease
Methyltransferases - genetics
Methyltransferases - metabolism
mRNA capping
Multienzyme Complexes - genetics
Nucleoside-Triphosphatase - metabolism
Nucleotidyltransferases - genetics
Nucleotidyltransferases - metabolism
Phosphoric Monoester Hydrolases - genetics
RNA, Messenger - metabolism
RNA, Viral - genetics
Transcription initiation
Transcription Initiation, Genetic - physiology
Transcription termination
Transcription Termination, Genetic - physiology
Vaccinia virus
Vaccinia virus - genetics
Vaccinia virus - metabolism
Viral Proteins
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Summary:Abstract Prior biochemical analysis of the heterodimeric vaccinia virus mRNA capping enzyme suggests roles not only in mRNA capping but also in early viral gene transcription termination and intermediate viral gene transcription initiation. Prior phenotypic characterization of Dts36, a temperature sensitive virus mutant affecting the large subunit of the capping enzyme was consistent with the multifunctional roles of the capping enzyme in vivo. We report a biochemical analysis of the capping enzyme encoded by Dts36. Of the three enzymatic activities required for mRNA capping, the guanylyltransferase and methyltransferase activities are compromised while the triphosphatase activity and the D12 subunit interaction are unaffected. The mutant enzyme is also defective in stimulating early gene transcription termination and intermediate gene transcription initiation in vitro. These results confirm that the vaccinia virus mRNA capping enzyme functions not only in mRNA capping but also early gene transcription termination and intermediate gene transcription initiation in vivo.
ISSN:0042-6822
1096-0341
DOI:10.1016/j.virol.2015.10.011