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

Authors

Phat X. Dinh, University of Nebraska-LincolnFollow
Debasis Panda, University of Nebraska - LincolnFollow
Phani B. Das, University of Nebraska-LincolnFollow
Subash C. Das, University of Nebraska - LincolnFollow
Anshuman Das, University of Nebraska-LincolnFollow
Asit K. Pattnaik, University of Nebraska-LincolnFollow

Document Type

Article

Date of this Version

2012

Citation

Virology 432 (2012) 460-469

Comments

© 2012 Elsevier Inc. All rights reserved.

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.