Association of hepatitis B virus polymerase with promyelocytic leukemia nuclear bodies mediated by the S100 family protein p11.
Hepatitis B virus (HBV) polymerase (Pol) interacts with cellular chaperone proteins and thereby performs multiple functions necessary for viral replication. Yeast two-hybrid analysis was applied to identify additional cellular targets required for HBV Pol function. HBV Pol interacted with S100A10 (p11), a Ca(2+)-modulated protein previously shown to bind to annexin II. The interaction between HBV Pol and p11 was confirmed by co-immunoprecipitation of the two proteins synthesized either in vitro or in transfected cells and by inhibition of the DNA polymerase activity of HBV Pol by p11. Immunofluorescence analysis of transfected human cell lines revealed that, although most HBV Pol and p11 was restricted to the cytoplasm, a small proportion of each protein colocalized as nuclear speckles; HBV Pol was not detected in the nucleus in the absence of p11. The HBV Pol-p11 nuclear speckles coincided with nuclear bodies containing the promyelocytic leukemia protein PML. Furthermore, the association of HBV Pol-p11 with PML was increased by exposure of cells to EGTA and inhibited by valinomycin. These results suggest a role for p11 in modulation of HBV Pol function and implicate PML nuclear bodies and intracellular Ca(2+) in viral replication.
Choi
Juhyun
J
Biomedical Research Center, Korea Institute of Science and Technology, 39-1 Hawallkok-dong, Sungbuk-ku, Seoul 136-791, South Korea.
Chang
Jin-Sook
JS
Song
Min-Sup
MS
Ahn
Byung-Yoon
BY
Park
Young
Y
Lim
Dae-Sik
DS
Han
Ye Sun
YS
D017306:Annexin A2; D002118:Calcium / Q000502:physiology; D002135:Calcium-Binding Proteins / Q000032:analysis / Q000378:metabolism*; D002467:Cell Nucleus / Q000737:chemistry* / Q000648:ultrastructure; D002614:Chelating Agents / Q000494:pharmacology; D003593:Cytoplasm / Q000737:chemistry; D004533:Egtazic Acid / Q000494:pharmacology; D015684:Gene Products, pol / Q000032:analysis* / Q000378:metabolism; D006801:Humans; D008856:Microscopy, Fluorescence; D009363:Neoplasm Proteins / Q000032:analysis*; D009687:Nuclear Proteins; D000071156:Promyelocytic Leukemia Protein; D009418:S100 Proteins; D014157:Transcription Factors / Q000032:analysis*; D014407:Tumor Cells, Cultured; D025521:Tumor Suppressor Proteins; D020798:Two-Hybrid System Techniques; D000099248:S100 Calcium Binding Protein A10
D017306:Annexin A2; D002118:Calcium; D002135:Calcium-Binding Proteins; D002614:Chelating Agents; D004533:Egtazic Acid; D015684:Gene Products, pol; D009363:Neoplasm Proteins; D009687:Nuclear Proteins; D000071156:Promyelocytic Leukemia Protein; D009418:S100 Proteins; D014157:Transcription Factors; D025521:Tumor Suppressor Proteins; D000099248:S100 Calcium Binding Protein A10; C066027:P protein, Hepatitis B virus; C070576:PML protein, human