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Title: A single amino acid change resulting in loss of fluorescence of eGFP in a viral fusion protein confers fitness and growth advantage to the recombinant vesicular stomatitis virus

Abstract

Using a recombinant vesicular stomatitis virus encoding eGFP fused in-frame with an essential viral replication protein, the phosphoprotein P, we show that during passage in culture, the virus mutates the nucleotide C289 within eGFP of the fusion protein PeGFP to A or T, resulting in R97S/C amino acid substitution and loss of fluorescence. The resultant non-fluorescent virus exhibits increased fitness and growth advantage over its fluorescent counterpart. The growth advantage of the non-fluorescent virus appears to be due to increased transcription and replication activities of the PeGFP protein carrying the R97S/C substitution. Further, our results show that the R97S/C mutation occurs prior to accumulation of mutations that can result in loss of expression of the gene inserted at the G-L gene junction. These results suggest that fitness gain is more important for the recombinant virus than elimination of expression of the heterologous gene.

Authors:
; ; ; ;  [1];  [2];  [1];  [2]
  1. School of Veterinary Medicine and Biomedical Sciences, University of Nebraska-Lincoln, Lincoln, Nebraska 68583-0900 (United States)
  2. (United States)
Publication Date:
OSTI Identifier:
22150074
Resource Type:
Journal Article
Journal Name:
Virology
Additional Journal Information:
Journal Volume: 432; Journal Issue: 2; Other Information: Copyright (c) 2012 Elsevier Science B.V., Amsterdam, The Netherlands, All rights reserved.; Country of input: International Atomic Energy Agency (IAEA); Journal ID: ISSN 0042-6822
Country of Publication:
United States
Language:
English
Subject:
60 APPLIED LIFE SCIENCES; AMINO ACIDS; FLUORESCENCE; MUTATIONS; NUCLEOTIDES; PHOSPHOPROTEINS; TRANSCRIPTION; VIRUSES

Citation Formats

Dinh, Phat X., Panda, Debasis, Das, Phani B., Das, Subash C., Das, Anshuman, The Nebraska Center for Virology, University of Nebraska-Lincoln, Lincoln, Nebraska 68583-0900, Pattnaik, Asit K., E-mail: apattnaik2@unl.edu, and The Nebraska Center for Virology, University of Nebraska-Lincoln, Lincoln, Nebraska 68583-0900. A single amino acid change resulting in loss of fluorescence of eGFP in a viral fusion protein confers fitness and growth advantage to the recombinant vesicular stomatitis virus. United States: N. p., 2012. Web. doi:10.1016/J.VIROL.2012.07.004.
Dinh, Phat X., Panda, Debasis, Das, Phani B., Das, Subash C., Das, Anshuman, The Nebraska Center for Virology, University of Nebraska-Lincoln, Lincoln, Nebraska 68583-0900, Pattnaik, Asit K., E-mail: apattnaik2@unl.edu, & The Nebraska Center for Virology, University of Nebraska-Lincoln, Lincoln, Nebraska 68583-0900. A single amino acid change resulting in loss of fluorescence of eGFP in a viral fusion protein confers fitness and growth advantage to the recombinant vesicular stomatitis virus. United States. doi:10.1016/J.VIROL.2012.07.004.
Dinh, Phat X., Panda, Debasis, Das, Phani B., Das, Subash C., Das, Anshuman, The Nebraska Center for Virology, University of Nebraska-Lincoln, Lincoln, Nebraska 68583-0900, Pattnaik, Asit K., E-mail: apattnaik2@unl.edu, and The Nebraska Center for Virology, University of Nebraska-Lincoln, Lincoln, Nebraska 68583-0900. Thu . "A single amino acid change resulting in loss of fluorescence of eGFP in a viral fusion protein confers fitness and growth advantage to the recombinant vesicular stomatitis virus". United States. doi:10.1016/J.VIROL.2012.07.004.
@article{osti_22150074,
title = {A single amino acid change resulting in loss of fluorescence of eGFP in a viral fusion protein confers fitness and growth advantage to the recombinant vesicular stomatitis virus},
author = {Dinh, Phat X. and Panda, Debasis and Das, Phani B. and Das, Subash C. and Das, Anshuman and The Nebraska Center for Virology, University of Nebraska-Lincoln, Lincoln, Nebraska 68583-0900 and Pattnaik, Asit K., E-mail: apattnaik2@unl.edu and The Nebraska Center for Virology, University of Nebraska-Lincoln, Lincoln, Nebraska 68583-0900},
abstractNote = {Using a recombinant vesicular stomatitis virus encoding eGFP fused in-frame with an essential viral replication protein, the phosphoprotein P, we show that during passage in culture, the virus mutates the nucleotide C289 within eGFP of the fusion protein PeGFP to A or T, resulting in R97S/C amino acid substitution and loss of fluorescence. The resultant non-fluorescent virus exhibits increased fitness and growth advantage over its fluorescent counterpart. The growth advantage of the non-fluorescent virus appears to be due to increased transcription and replication activities of the PeGFP protein carrying the R97S/C substitution. Further, our results show that the R97S/C mutation occurs prior to accumulation of mutations that can result in loss of expression of the gene inserted at the G-L gene junction. These results suggest that fitness gain is more important for the recombinant virus than elimination of expression of the heterologous gene.},
doi = {10.1016/J.VIROL.2012.07.004},
journal = {Virology},
issn = {0042-6822},
number = 2,
volume = 432,
place = {United States},
year = {2012},
month = {10}
}