5' cis elements direct nodavirus RNA1 recruitment to mitochondrial sites of replication complex formation.

Journal of Virology
Priscilla M Van Wynsberghe, Paul Ahlquist

Abstract

Positive-strand RNA viruses replicate their genomes on intracellular membranes, usually in conjunction with virus-induced membrane rearrangements. For the nodavirus flock house virus (FHV), we recently showed that multifunctional FHV replicase protein A induces viral RNA template recruitment to a membrane-associated state, but the site(s) and function of this recruitment were not determined. By tagging viral RNA with green fluorescent protein, we show here in Drosophila cells that protein A recruits FHV RNA specifically to the outer mitochondrial membrane sites of RNA replication complex formation. Using Drosophila cells and yeast cells, which also support FHV replication, we also defined the cis-acting regions that direct replication and template recruitment for FHV genomic RNA1. RNA1 nucleotides 68 to 205 were required for RNA replication and directed efficient protein A-mediated RNA recruitment in both cell types. RNA secondary structure prediction, structure probing, and phylogenetic comparisons in this region identified two stable, conserved stem-loops with nearly identical loop sequences. Further mutational analysis showed that both stem-loops and certain flanking sequences were required for RNA1 recruitment, negative-str...Continue Reading

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Citations

Dec 21, 2010·Apoptosis : an International Journal on Programmed Cell Death·Yu-Chin SuJiann-Ruey Hong
Sep 10, 2010·Annual Review of Microbiology·Johan A den Boon, Paul Ahlquist
Sep 29, 2009·Biochimica Et Biophysica Acta·Ying LiuAniko V Paul
Sep 27, 2015·Methods : a Companion to Methods in Enzymology·Andrew RouthBruce E Torbett
Mar 22, 2013·The Journal of General Virology·Suzanne U EmersonAndrew E Firth
Apr 21, 2011·RNA Biology·Zhenghe Li, Peter D Nagy
Jul 20, 2010·Cell Host & Microbe·Johan A den BoonPaul Ahlquist
Dec 21, 2011·Nature Reviews. Microbiology·Peter D Nagy, Judit Pogany

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