Involvement of Crm1 in hepatitis B virus X protein-induced aberrant centriole replication and abnormal mitotic spindles.
Hepatitis B virus (HBV) includes an X gene (HBx gene) that plays a critical role in liver carcinogenesis. Because centrosome abnormalities are associated with genomic instability in most human cancer cells, we examined the effect of HBx on centrosomes. We found that HBx induced supernumerary centrosomes and multipolar spindles. This effect was independent of mutations in the p21 gene. Furthermore, the ability of HBV to induce supernumerary centrosomes was dependent on the presence of physiological HBx expression. We recently showed that HBx induces cytoplasmic sequestration of Crm1, a nuclear export receptor that binds to Ran GTPase, thereby inducing nuclear localization of NF-kappaB. Consistently, supernumerary centrosomes were observed in cells treated with a Crm1-specific inhibitor but not with an HBx mutant that lacked the ability to sequester Crm1 in the cytoplasm. Moreover, a fraction of Crm1 was found to be localized at the centrosomes. Immunocytochemical and ultrastructural examination of these supernumerary centrosomes revealed that inactivation of Crm1 was associated with abnormal centrioles. The presence of more than two centrosomes led to an increased frequency of defective mitoses and chromosome transmission errors. Based on this evidence, we suggest that Crm1 is actively involved in maintaining centrosome integrity and that HBx disrupts this process by inactivating Crm1 and thus contributes to HBV-mediated carcinogenesis.
Forgues
Marshonna
M
Laboratory of Human Carcinogenesis, Center for Cancer Research, National Cancer Institute/NIH, 37 Convent Drive, Bethesda, MD 20892, USA.
Difilippantonio
Michael J
MJ
Linke
Steven P
SP
Ried
Thomas
T
Nagashima
Kunio
K
Feden
Jeffrey
J
Valerie
Kristoffer
K
Fukasawa
Kenji
K
Wang
Xin W
XW
D021581:Active Transport, Cell Nucleus; D000256:Adenoviridae / Q000235:genetics; D000782:Aneuploidy; D015153:Blotting, Western; D002453:Cell Cycle; D002467:Cell Nucleus / Q000378:metabolism; D002502:Centrioles / Q000378:metabolism*; D018385:Centrosome; D003593:Cytoplasm / Q000378:metabolism; D004247:DNA / Q000378:metabolism; D004268:DNA-Binding Proteins; D005231:Fatty Acids, Unsaturated / Q000494:pharmacology; D005347:Fibroblasts / Q000378:metabolism; D019084:Fluorescent Antibody Technique, Indirect; D006515:Hepatitis B virus / Q000378:metabolism; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D028884:Karyopherins / Q000502:physiology*; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D008938:Mitosis; D009154:Mutation; D016328:NF-kappa B / Q000378:metabolism; D018160:Receptors, Cytoplasmic and Nuclear; D008941:Spindle Apparatus; D019098:Telomerase / Q000378:metabolism; D013997:Time Factors; D015534:Trans-Activators / Q000378:metabolism*; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism; D054334:Viral Regulatory and Accessory Proteins; D020931:ran GTP-Binding Protein / Q000378:metabolism; D020741:rho GTP-Binding Proteins / Q000378:metabolism; D000097606:Exportin 1 Protein
D004247:DNA; D004268:DNA-Binding Proteins; D005231:Fatty Acids, Unsaturated; D028884:Karyopherins; D016328:NF-kappa B; D018160:Receptors, Cytoplasmic and Nuclear; D019098:Telomerase; D015534:Trans-Activators; D016159:Tumor Suppressor Protein p53; D054334:Viral Regulatory and Accessory Proteins; D020931:ran GTP-Binding Protein; D020741:rho GTP-Binding Proteins; D000097606:Exportin 1 Protein; C057628:hepatitis B virus X protein; C038753:leptomycin B