title: A Homokaryon Assay for Nucleocytoplasmic Shuttling Activity of HBV Core Protein.
abstract: Hepatitis B virus (HBV) core protein (HBc) can be present in both nucleus and cytoplasm. The arginine-rich domain (ARD) at the cytoplasmic tail of HBc contains both a nuclear localization signal (NLS) and nuclear export signal (NES). We established a homokaryon assay to detect the dynamic trafficking of HBc between nucleus and cytoplasm in hepatocytes. Using immunofluorescence assay (IFA) and PEG-induced cell-cell fusion, we demonstrated that a chimeric reporter protein of SV40 large T antigen, when fused in-frame with HBc ARD, can shuttle from a donor nuclei (green) to the recipient nuclei (red) in the context of binucleated or polynucleated hybrid cells. The shuttling activity driven by HBc ARD can be measured quantitatively by this IFA method.
authors.affiliation: Taiwan International Graduate Program (TIGP) in Molecular Medicine, National Yang-Ming University and Academia Sinica, Taipei, Taiwan.|Institute of Biomedical Sciences, Academia Sinica, Taipei, Taiwan.
authors.forename: Ching-Chun
authors.initials: CC
authors.lastname: Yang
authors.affiliation: Institute of Biomedical Sciences, Academia Sinica, Taipei, Taiwan.
authors.forename: Hung-Cheng
authors.initials: HC
authors.lastname: Li
authors.affiliation: Institute of Biomedical Sciences, Academia Sinica, Taipei, Taiwan. cshih@ibms.sinica.edu.
authors.forename: Chiaho
authors.initials: C
authors.lastname: Shih
mesh_terms: D045744:Cell Line, Tumor; D002467:Cell Nucleus / Q000378:metabolism; D003593:Cytoplasm / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D006509:Hepatitis B / Q000821:virology*; D006515:Hepatitis B virus / Q000502:physiology*; D006801:Humans; D054730:Protein Interaction Domains and Motifs; D021381:Protein Transport; D014758:Viral Core Proteins / Q000737:chemistry / Q000378:metabolism*
chemical_list: D014758:Viral Core Proteins
keywords: Arginine-rich domain (ARD); HBV core; Nuclear export; Nuclear import; Nucleocytoplasmic shuttling