ORF3 protein of hepatitis E virus is not required for replication, virion assembly, or infection of hepatoma cells in vitro.
A subclone of Huh-7 cells that could be relatively efficiently transfected and infected with hepatitis E virus was identified. Following transfection, infectious virus was produced but remained predominantly cell associated. Intracellular virus, recovered by lysis of transfected cells, infected naïve cells. This in vitro-produced virus appeared to be antigenically identical to virus isolated from clinical samples. Lysates from cells transfected with mutant viral genomes unable to synthesize ORF3 protein contained infectious virions that were similar in number, thermostability, and sedimentation characteristics to those in lysates transfected with wild-type viral genomes. Therefore, in contrast to its requirement in vivo, ORF3 protein is not required for infection of Huh-7 cells or production of infectious virus in vitro.
Emerson
Suzanne U
SU
Laboratory of Infectious Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Building 50, Room 6537, 50 South Drive, MSC 8009, Bethesda, MD 20892, USA. semerson@niaid.nih.gov
Nguyen
Hanh
H
Torian
Udana
U
Purcell
Robert H
RH
D006528:Carcinoma, Hepatocellular / Q000821:virology; D045744:Cell Line, Tumor; D016752:Hepatitis E virus / Q000235:genetics / Q000472:pathogenicity / Q000502:physiology*; D006801:Humans; D066298:In Vitro Techniques; D009500:Neutralization Tests; D014162:Transfection; D014764:Viral Proteins / Q000235:genetics / Q000502:physiology*; D014774:Virulence; D019065:Virus Assembly; D014779:Virus Replication
C400971:ORF2 protein, Hepatitis E virus; C400973:ORF3 protein, Hepatitis E virus; D014764:Viral Proteins