Viable polioviruses that encode 2A proteins with fluorescent protein tags.

Journal of Virology
N TeterinaE Ehrenfeld

Abstract

The 2A proteins of the Picornaviridae enterovirus genus are small cysteine proteinases that catalyze essential cleavages in the viral polyprotein in cis and in several cellular proteins in trans. In addition, 2A has been implicated in the process of viral RNA replication, independent of its protease functions. We have generated viable polioviruses that encode 2A proteins containing fluorescent protein tag insertions at either of two sites in the 2A protein structure. Viruses containing an insertion of Discosoma sp. red fluorescent protein (DsRed) after residue 144 of 2A, near the C terminus, produced plaques only slightly smaller than wild-type (wt) virus. The polyprotein harboring the 2A-DsRed fusion protein was efficiently and accurately cleaved; fluorescent 2A proteinase retained protease activity in trans and supported translation and replication of viral RNA, both in vitro and in infected cells. Intracellular membrane reorganization to support viral RNA synthesis was indistinguishable from that induced by wt virus. Infected cells exhibited strong red fluorescence from expression of the 2A-DsRed fusion protein, and the progeny virus was stable for three to four passages, after which deletions within the DsRed coding sequenc...Continue Reading

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Citations

Aug 24, 2012·Journal of Virology·Lucy ThorneIan Goodfellow
Feb 25, 2011·Journal of Virology·Natalya L TeterinaEllie Ehrenfeld
Dec 14, 2011·Journal of Virology·Yutong SongEckard Wimmer
May 5, 2011·Journal of Biomedicine & Biotechnology·Alfredo CastellóLuis Carrasco
Aug 27, 2014·Proceedings of the National Academy of Sciences of the United States of America·Sara Whitney BirdKarla Kirkegaard
Oct 4, 2015·The Journal of General Virology·Morgan R HerodNicola J Stonehouse
Mar 11, 2016·Cell Host & Microbe·Troels K H ScheelCharles M Rice
Oct 26, 2010·Virology·Natalya L TeterinaEllie Ehrenfeld
Apr 22, 2015·Virology·Sara W Bird, Karla Kirkegaard
Dec 14, 2018·Journal of Virology·Oluwapelumi O AdeyemiNicola J Stonehouse
Sep 22, 2019·Viruses·Claire MuslinFrancis Delpeyroux
Mar 16, 2018·Microbiology and Molecular Biology Reviews : MMBR·Vadim I Agol, Anatoly P Gmyl
Jan 13, 2017·Nature·Jacqueline StaringThijn R Brummelkamp
Aug 9, 2017·Frontiers in Microbiology·Xiaoyao YangXiaoyue Chen

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