Repository logo

Characterization of the stimulators of protein-directed ribosomal frameshifting in Theiler's murine encephalomyelitis virus.

Accepted version

Change log


Bell, Susanne 
Hill, Chris H 
Firth, Andrew E 


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.



Animals, Base Sequence, Frameshifting, Ribosomal, Mice, Nucleic Acid Conformation, Protein Biosynthesis, RNA, Messenger, RNA, Viral, Ribosomes, Theilovirus, Viral Proteins

Journal Title

Nucleic Acids Res

Conference Name

Journal ISSN


Volume Title



Oxford University Press (OUP)


All rights reserved
Biotechnology and Biological Sciences Research Council (BB/J007072/1)
Wellcome Trust (106207/Z/14/Z)
European Research Council (646891)
Biotechnology and Biological Sciences Research Council (BB/L000334/1)
Medical Research Council (MR/M011747/1)
Wellcome Trust (202797/Z/16/Z)