Napthine, Sawsan, Bell, Susanne, Hill, Chris H orcid.org/0000-0001-7037-0611 et al. (2 more authors) (2019) Characterization of the stimulators of protein-directed ribosomal frameshifting in Theiler's murine encephalomyelitis virus. Nucleic Acids Research. pp. 8207-8223. ISSN 0305-1048
Abstract
Many viruses utilize programmed -1 ribosomal frameshifting (-1 PRF) to express additional proteins or to produce frameshift and non-frameshift protein products at a fixed stoichiometric ratio. PRF is also utilized in the expression of a small number of cellular genes. Frameshifting is typically stimulated by signals contained within the mRNA: a 'slippery' sequence and a 3'-adjacent RNA structure. Recently, we showed that -1 PRF in encephalomyocarditis virus (EMCV) is trans-activated by the viral 2A protein, leading to a temporal change in PRF efficiency from 0% to 70% during virus infection. Here we analyzed PRF in the related Theiler's murine encephalomyelitis virus (TMEV). We show that 2A is also required for PRF in TMEV and can stimulate PRF to levels as high as 58% in rabbit reticulocyte cell-free translations and 81% during virus infection. We also show that TMEV 2A trans-activates PRF on the EMCV signal but not vice versa. We present an extensive mutational analysis of the frameshift stimulators (mRNA signals and 2A protein) analysing activity in in vitro translation, electrophoretic mobility shift and in vitro ribosome pausing assays. We also investigate the PRF mRNA signal with RNA structure probing. Our results substantially extend previous characterization of protein-stimulated PRF.
Metadata
Item Type: | Article |
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Authors/Creators: |
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Copyright, Publisher and Additional Information: | © The Author(s) 2019. Published by Oxford University Press on behalf of Nucleic Acids Research. |
Keywords: | Animals,Base Sequence,Frameshifting, Ribosomal/genetics,Mice,Nucleic Acid Conformation,Protein Biosynthesis/genetics,RNA, Messenger/chemistry,RNA, Viral/chemistry,Ribosomes/genetics,Theilovirus/genetics,Viral Proteins/genetics |
Dates: |
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Institution: | The University of York |
Academic Units: | The University of York > Faculty of Sciences (York) > Biology (York) |
Depositing User: | Pure (York) |
Date Deposited: | 22 Oct 2021 11:00 |
Last Modified: | 24 Oct 2024 00:14 |
Published Version: | https://doi.org/10.1093/nar/gkz503 |
Status: | Published |
Refereed: | Yes |
Identification Number: | 10.1093/nar/gkz503 |
Open Archives Initiative ID (OAI ID): | oai:eprints.whiterose.ac.uk:179528 |
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Description: Characterization of the stimulators of protein-directed ribosomal frameshifting in Theiler’s murine encephalomyelitis virus
Licence: CC-BY 2.5