RRC ID 51804
Author Nagashima S, Kobayashi T, Tanaka T, Tanggis, Jirintai S, Takahashi M, Nishizawa T, Okamoto H.
Title Analysis of adaptive mutations selected during the consecutive passages of hepatitis E virus produced from an infectious cDNA clone.
Journal Virus Res
Abstract To characterize the genomic mutations of hepatitis E virus (HEV) during consecutive passages associated with adaptation to growth in cell culture, a cloned genotype 3 HEV [pJE03-1760F/wt, starting virus (SV)] was passaged 10 times in A549 cells, and the entire genomic sequence of the passage 10 (P10) progeny was determined. Compared to SV, P10 virus possessed two non-synonymous (T2808C and A5054G) and four synonymous mutations (C1213T, T2557C, C3118T and C4435T) in the ORF1. Full-length infectious cDNA clones with a single, double (T2808C and A5054G), or all six mutations, identical to P10, were constructed, and their replication capacity was compared. Four (C1213T, T2557C, T2808C and A5054G) of the six viruses with a single mutation grew more efficiently than SV. The P10 virus propagated more rapidly and grew more efficiently than SV and T2808C+A5054G and reached a higher viral load (95.1- and 8.5-fold, respectively) at 20days post-inoculation. An immunofluorescence analysis revealed that a high percentage (>80%) of cells inoculated with the P10 virus expressed ORF2 proteins, while relatively low percentages (nearly 30% or 5%) inoculated with T2808C+A5054G or SV, respectively, expressed ORF2 proteins. We found that not only non-synonymous but also synonymous HEV mutations are independently associated with increased virus production.
Volume 223
Pages 170-80
Published 2016-9-2
DOI 10.1016/j.virusres.2016.07.011
PII S0168-1702(16)30391-4
PMID 27485920
MeSH Adaptation, Biological / genetics* Amino Acid Substitution Cell Line DNA, Complementary Genome, Viral Hepatitis E virus / classification Hepatitis E virus / genetics* Humans Mutation* Phylogeny RNA, Viral* Reassortant Viruses / genetics Recombination, Genetic Selection, Genetic* Viral Load Virus Cultivation Virus Replication
IF 2.934
Times Cited 2
Human and Animal Cells A549(RCB0098)