Mutational analysis of hepatitis E virus ORF1 "Y-domain": Effects on RNA replication and virion infectivity.

ABSTRACT:

Aim

To investigate the role of non-structural open reading frame 1 "Y-domain" sequences in the hepatitis E virus (HEV) life cycle.

Methods

Sequences of human HEV Y-domain (amino acid sequences 216-442) and closely-related viruses were analyzed in silico. Site-directed mutagenesis of the Y-domain (HEV SAR55) was carried out and studied in the replicon-baculovirus-hepatoma cell model. In vitro transcribed mRNA (pSK-GFP) constructs were transfected into S10-3 cells and viral RNA replicating GFP-positive cells were scored by flow cytometry. Mutant virions' infectivity was assayed on naïve HepG2/C3A cells.

Results

In silico analysis identified a potential palmitoylation-site (C₃₃₆C₃₃₇) and an α-helix segment (L₄₁₀

SUBMITTER: Parvez MK 

PROVIDER: S-EPMC5292332 | biostudies-literature | 2017 Jan

REPOSITORIES: biostudies-literature