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Modulation of Viral Programmed Ribosomal Frameshifting and Stop Codon Readthrough by the Host Restriction Factor Shiftless.

Published version
Peer-reviewed

Type

Article

Change log

Authors

Hill, Chris H 
Nugent, Holly CM 

Abstract

The product of the interferon-stimulated gene C19orf66, Shiftless (SHFL), restricts human immunodeficiency virus replication through downregulation of the efficiency of the viral gag/pol frameshifting signal. In this study, we demonstrate that bacterially expressed, purified SHFL can decrease the efficiency of programmed ribosomal frameshifting in vitro at a variety of sites, including the RNA pseudoknot-dependent signals of the coronaviruses IBV, SARS-CoV and SARS-CoV-2, and the protein-dependent stimulators of the cardioviruses EMCV and TMEV. SHFL also reduced the efficiency of stop-codon readthrough at the murine leukemia virus gag/pol signal. Using size-exclusion chromatography, we confirm the binding of the purified protein to mammalian ribosomes in vitro. Finally, through electrophoretic mobility shift assays and mutational analysis, we show that expressed SHFL has strong RNA binding activity that is necessary for full activity in the inhibition of frameshifting, but shows no clear specificity for stimulatory RNA structures.

Description

Keywords

Shiftless, frameshift, readthrough, ribosome, virus, Base Sequence, Codon, Terminator, Coronavirus, Escherichia coli, Frameshifting, Ribosomal, Gene Expression Regulation, Viral, Humans, Leukemia Virus, Murine, RNA Recognition Motif Proteins, RNA, Viral, RNA-Binding Proteins, Viral Proteins, Virus Replication

Journal Title

Viruses

Conference Name

Journal ISSN

1999-4915
1999-4915

Volume Title

13

Publisher

MDPI AG
Sponsorship
Wellcome Trust (202797/Z/16/Z)
BBSRC (BB/V000306/1)