title: A Semliki Forest virus expression system as a model for investigating the nuclear import and export of hepatitis B virus nucleocapsid protein.
abstract: HBV core protein (HBc), the major component of nucleocapsid is known to have an essential role in the virus life cycle as the transporter of virus genome to the host nucleus; however, molecular details of the intracellular transport of HBc remain unknown. In this study, we investigated the intracellular distribution of the HBc protein resulting from Semliki Forest virus (SFV)-driven HBc gene and HBV RNA pregenome (pgRNA) expression in BHK-21 cells. Fluorescent confocal microscopy revealed extensive HBc protein synthesis in the cytoplasm 12 hr post infection (p.i.) with recombinant pSFV1/HBc or pSFV1/HBVpgRNA viruses. Twenty-four hr p.i., the HBc protein showed asymmetric, predominantly nuclear localization in most cells. However, 42 hr p.i., the number of cells containing intranuclear HBc protein, dramatically decreased to ~ 5%, while cytoplasmic HBc protein was detected in all cells. Remarkably, 24 hr p.i. with pSFV1/HBVpgRNA virus, cytoplasmic HBc protein colocalized with HBs protein. We conclude that a HBs-HBc interaction partially prevents the transport of HBc from the cytoplasm to nucleus. The SFV-driven system can be used for identification of new factors involved in the HBV nuclear import and export.
authors: A
authors: A
authors: Zajakina
authors: R
authors: R
authors: Brūvere
authors: T
authors: T
authors: Kozlovska
mesh_terms: D021581:Active Transport, Cell Nucleus; D001692:Biological Transport; D002467:Cell Nucleus / Q000378:metabolism*; D003593:Cytoplasm / Q000378:metabolism; D006509:Hepatitis B / Q000378:metabolism / Q000821:virology*; D006515:Hepatitis B virus / Q000378:metabolism*; D006801:Humans; D019590:Nucleocapsid Proteins / Q000378:metabolism*; D012672:Semliki forest virus / Q000235:genetics / Q000378:metabolism*
chemical_list: D019590:Nucleocapsid Proteins; C099600:nucleocapsid protein, Hepatitis virus