Open AccessIdentification of GBF1 as a cellular factor required for hepatitis E virus RNA replication
Author(s) -
Farhat Rayan,
Ankavay Maliki,
Lebsir Nadjet,
Gouttenoire Jérôme,
Jackson Catherine L.,
Wychowski Czeslaw,
Moradpour Darius,
Dubuisson Jean,
Rouillé Yves,
Cocquerel Laurence
Publication year - 2018
Publication title -
cellular microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.542
H-Index - 138
eISSN - 1462-5822
pISSN - 1462-5814
DOI - 10.1111/cmi.12804
Subject(s) - biology , host factor , viral replication , virology , replication factor c , guanine nucleotide exchange factor , hepatitis e virus , rna , microbiology and biotechnology , virus , dna replication , gtpase , genetics , control of chromosome duplication , dna , gene , genotype
Abstract The hepatitis E virus (HEV) genome is a single‐stranded, positive‐sense RNA that encodes three proteins including the ORF1 replicase. Mechanisms of HEV replication in host cells are unclear, and only a few cellular factors involved in this step have been identified so far. Here, we used brefeldin A (BFA) that blocks the activity of the cellular Arf guanine nucleotide exchange factors GBF1, BIG1, and BIG2, which play a major role in reshuffling of cellular membranes. We showed that BFA inhibits HEV replication in a dose‐dependent manner. The use of siRNA and Golgicide A identified GBF1 as a host factor critically involved in HEV replication. Experiments using cells expressing a mutation in the catalytic domain of GBF1 and overexpression of wild type GBF1 or a BFA‐resistant GBF1 mutant rescuing HEV replication in BFA‐treated cells, confirmed that GBF1 is the only BFA‐sensitive factor required for HEV replication. We demonstrated that GBF1 is likely required for the activity of HEV replication complexes. However, GBF1 does not colocalise with the ORF1 protein, and its subcellular distribution is unmodified upon infection or overexpression of viral proteins, indicating that GBF1 is likely not recruited to replication sites. Together, our results suggest that HEV replication involves GBF1‐regulated mechanisms.