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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 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.
AuthorsJessica Tate, Rachel L Boldt, Baron D McFadden, Susan M D'Costa, Nicholas M Lewandowski, Amber N Shatzer, Paul Gollnick, Richard C Condit
JournalVirology (Virology) Vol. 487 Pg. 27-40 (Jan 2016) ISSN: 1096-0341 [Electronic] United States
PMID26496697 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, U.S. Gov't, Non-P.H.S.)
CopyrightCopyright © 2015 Elsevier Inc. All rights reserved.
Chemical References
  • Multienzyme Complexes
  • RNA, Messenger
  • RNA, Viral
  • Viral Proteins
  • Methyltransferases
  • Nucleotidyltransferases
  • mRNA guanylyltransferase
  • Phosphoric Monoester Hydrolases
  • capping enzyme, vaccinia virus
  • Nucleoside-Triphosphatase
Topics
  • Animals
  • Cell Line
  • Chlorocebus aethiops
  • HeLa Cells
  • Humans
  • Methyltransferases (genetics, metabolism)
  • Multienzyme Complexes (genetics)
  • Nucleoside-Triphosphatase (metabolism)
  • Nucleotidyltransferases (genetics, metabolism)
  • Phosphoric Monoester Hydrolases (genetics)
  • RNA, Messenger (metabolism)
  • RNA, Viral (genetics)
  • Transcription Initiation, Genetic (physiology)
  • Transcription Termination, Genetic (physiology)
  • Vaccinia virus (genetics, metabolism)
  • Viral Proteins

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