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: School of Veterinary Medicine and Biomedical Sciences, University of Nebraska-Lincoln, Lincoln, Nebraska 68583-0900, USA.
authors: Phat X
authors: PX
authors: Dinh
authors: Debasis
authors: D
authors: Panda
authors: Phani B
authors: PB
authors: Das
authors: Subash C
authors: SC
authors: Das
authors: Anshuman
authors: A
authors: Das
authors: Asit K
authors: AK
authors: Pattnaik
mesh_terms: D019943:Amino Acid Substitution; D000818:Animals; D002460:Cell Line; D006224:Cricetinae; D005453:Fluorescence; D005456:Fluorescent Dyes; D049452:Green Fluorescent Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D011995:Recombination, Genetic; D014158:Transcription, Genetic; D014721:Vesicular stomatitis Indiana virus / Q000235:genetics / Q000254:growth & development* / Q000378:metabolism; D014760:Viral Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D014779:Virus Replication
chemical_list: D005456:Fluorescent Dyes; D014760:Viral Fusion Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins