Release of Genotype 1 Hepatitis E Virus from Cultured Hepatoma and Polarized Intestinal Cells Depends on Open Reading Frame 3 Protein and Requires an Intact PXXP Motif

ABSTRACT Hepatitis E virus genotype 1 strain Sar55 replicated in subcloned Caco-2 intestinal cells and Huh7 hepatoma cells that had been transfected with in vitro transcribed viral genomes, and hepatitis E virions were released into the culture medium of both cell lines. Virus egress from cells depended on open reading frame 3 (ORF3) protein, and a proline-rich sequence in ORF3 was important for egress from cultured cells and for infection of macaques. Both intracellular ORF3 protein accumulation and virus release occurred at the apical membrane of polarized Caco-2 cells. ORF3 protein and lipids were intimately associated with virus particles produced in either cell line; ORF2 epitopes were masked in these particles and could not be immunoprecipitated with anti-ORF2.

[1]  X. Meng,et al.  Recent advances in Hepatitis E Virus , 2010, Journal of viral hepatitis.

[2]  S. Jameel,et al.  The hepatitis E virus ORF3 protein stabilizes HIF‐1α and enhances HIF‐1‐mediated transcriptional activity through p300/CBP , 2009, Cellular microbiology.

[3]  Y. Tao,et al.  Structure of the hepatitis E virus-like particle suggests mechanisms for virus assembly and receptor binding , 2009, Proceedings of the National Academy of Sciences.

[4]  T. Tsukihara,et al.  Biological and immunological characteristics of hepatitis E virus-like particles based on the crystal structure , 2009, Proceedings of the National Academy of Sciences.

[5]  Toshinori Tanaka,et al.  ORF3 protein of hepatitis E virus is essential for virion release from infected cells. , 2009, The Journal of general virology.

[6]  J. Seetharaman,et al.  Dimerization of Hepatitis E Virus Capsid Protein E2s Domain Is Essential for Virus–Host Interaction , 2009, PLoS pathogens.

[7]  Youchun Wang,et al.  A novel genotype of hepatitis E virus prevalent among farmed rabbits in China , 2009, Journal of medical virology.

[8]  H. Mizuo,et al.  Development and Characterization of a Genotype 4 Hepatitis E Virus Cell Culture System Using a HE-JF5/15F Strain Recovered from a Fulminant Hepatitis Patient , 2009, Journal of Clinical Microbiology.

[9]  I. Bossis,et al.  The Hepatitis E Virus Open Reading Frame 3 Product Interacts with Microtubules and Interferes with Their Dynamics , 2009, Journal of Virology.

[10]  Toshinori Tanaka,et al.  Monoclonal antibodies raised against the ORF3 protein of hepatitis E virus (HEV) can capture HEV particles in culture supernatant and serum but not those in feces , 2008, Archives of Virology.

[11]  T. LeRoith,et al.  Development and Validation of a Negative-Strand-Specific Reverse Transcription-PCR Assay for Detection of a Chicken Strain of Hepatitis E Virus: Identification of Nonliver Replication Sites , 2008, Journal of Clinical Microbiology.

[12]  Sudha Kumari,et al.  The Hepatitis E Virus ORF3 Protein Modulates Epidermal Growth Factor Receptor Trafficking, STAT3 Translocation, and the Acute-Phase Response , 2008, Journal of Virology.

[13]  L. Xing,et al.  Crystallization and preliminary X-ray diffraction analysis of recombinant hepatitis E virus-like particle. , 2008, Acta crystallographica. Section F, Structural biology and crystallization communications.

[14]  A. Kar-Roy,et al.  The ORF3 protein of hepatitis E virus interacts with hemopexin by means of its 26 amino acid N-terminal hydrophobic domain II. , 2008, Biochemistry.

[15]  Syed M. Moin,et al.  The Hepatitis E Virus Orf3 Protein Protects Cells from Mitochondrial Depolarization and Death* , 2007, Journal of Biological Chemistry.

[16]  E. Kusano,et al.  Development and evaluation of an efficient cell-culture system for Hepatitis E virus. , 2007, The Journal of general virology.

[17]  T. Opriessnig,et al.  Initiation at the Third In-Frame AUG Codon of Open Reading Frame 3 of the Hepatitis E Virus Is Essential for Viral Infectivity In Vivo , 2007, Journal of Virology.

[18]  S. Emerson,et al.  ORF3 Protein of Hepatitis E Virus Is Not Required for Replication, Virion Assembly, or Infection of Hepatoma Cells In Vitro , 2006, Journal of Virology.

[19]  S. Emerson,et al.  A Bicistronic Subgenomic mRNA Encodes both the ORF2 and ORF3 Proteins of Hepatitis E Virus , 2006, Journal of Virology.

[20]  Milan Surjit,et al.  The 41-Amino-Acid C-Terminal Region of the Hepatitis E Virus ORF3 Protein Interacts with Bikunin, a Kunitz-Type Serine Protease Inhibitor , 2005, Journal of Virology.

[21]  M. St. Claire,et al.  The Open Reading Frame 3 Gene of Hepatitis E Virus Contains a cis-Reactive Element and Encodes a Protein Required for Infection of Macaques , 2005, Journal of Virology.

[22]  S. Emerson,et al.  In Vitro Replication of Hepatitis E Virus (HEV) Genomes and of an HEV Replicon Expressing Green Fluorescent Protein , 2004, Journal of Virology.

[23]  S. Mishiro,et al.  Zoonotic transmission of hepatitis E virus from deer to human beings , 2003, The Lancet.

[24]  S. Jameel,et al.  The Phosphorylated Form of the ORF3 Protein of Hepatitis E Virus Interacts with Its Non-glycosylated Form of the Major Capsid Protein, ORF2* , 2002, The Journal of Biological Chemistry.

[25]  Michael Ruogu Zhang,et al.  Recombinant hepatitis E virus genomes infectious for primates: Importance of capping and discovery of a cis-reactive element , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[26]  D. Gupta,et al.  The ORF3 Protein of Hepatitis E Virus Binds to Src Homology 3 Domains and Activates MAPK* , 2001, The Journal of Biological Chemistry.

[27]  P. Halbur,et al.  Evidence of Extrahepatic Sites of Replication of the Hepatitis E Virus in a Swine Model , 2001, Journal of Clinical Microbiology.

[28]  S. Lemon,et al.  Infection of Polarized Cultures of Human Intestinal Epithelial Cells with Hepatitis A Virus: Vectorial Release of Progeny Virions through Apical Cellular Membranes , 2000, Journal of Virology.

[29]  Division on Earth.  Guide for the Care and Use of Laboratory Animals , 1996 .

[30]  T. Yamane,et al.  Growth of human hepatoma cells lines with differentiated functions in chemically defined medium. , 1982, Cancer research.

[31]  J. Maniloff,et al.  Virus taxonomy : eighth report of the International Committee on Taxonomy of Viruses , 2005 .