Temporal Regulation of Distinct Internal Ribosome Entry Sites of the Dicistroviridae Cricket Paralysis Virus

Viruses
Anthony KhongEric Jan

Abstract

Internal ribosome entry is a key mechanism for viral protein synthesis in a subset of RNA viruses. Cricket paralysis virus (CrPV), a member of Dicistroviridae, has a positive-sense single strand RNA genome that contains two internal ribosome entry sites (IRES), a 5'untranslated region (5'UTR) and intergenic region (IGR) IRES, that direct translation of open reading frames (ORF) encoding the viral non-structural and structural proteins, respectively. The regulation of and the significance of the CrPV IRESs during infection are not fully understood. In this study, using a series of biochemical assays including radioactive-pulse labelling, reporter RNA assays and ribosome profiling, we demonstrate that while 5'UTR IRES translational activity is constant throughout infection, IGR IRES translation is delayed and then stimulated two to three hours post infection. The delay in IGR IRES translation is not affected by inhibiting global translation prematurely via treatment with Pateamine A. Using a CrPV replicon that uncouples viral translation and replication, we show that the increase in IGR IRES translation is dependent on expression of non-structural proteins and is greatly stimulated when replication is active. Temporal regulation ...Continue Reading

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Citations

Oct 30, 2016·RNA Biology·Samuel E Butcher, Eric Jan
Apr 4, 2017·The Journal of General Virology·S M Valles Ictv Report Consortium
Apr 8, 2016·Journal of Virology·Craig H Kerr, Eric Jan
Apr 22, 2017·Frontiers in Plant Science·Manuel MirasMiguel A Aranda
Dec 2, 2017·Wiley Interdisciplinary Reviews. RNA·Justine Mailliot, Franck Martin

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Methods Mentioned

BETA
transfection
fluorescence-focusing
immunoprecipitation
PCR
in vitro transcription
RNAseq
pulldowns
transfections

Software Mentioned

SOLiD
XSQ Tools
Snapdragon
ImageJ
ImageQuant

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