Production of hepatitis B virus nucleocapsidlike core particles in Xenopus oocytes: assembly occurs mainly in the cytoplasm and does not require the nucleus.
The location of hepatitis B virus (HBV) nucleocapsid (core particle) assembly in infected cells remains controversial. Some lines of evidence implicate the nucleus; others favor the cytoplasm. Via injection of a synthetic mRNA encoding the HBV nucleocapsid protein (p21.5), we have expressed both unassembled p21.5 and nucleocapsidlike core particles in Xenopus oocytes. Subcellular fractionation reveals that approximately 91% of the unassembled p21.5 and 95% of the core particles are cytoplasmic, with only 9 and 5%, respectively, in the nucleus. We present evidence showing that unassembled p21.5 equilibrates between nucleus and cytoplasm by passive diffusion and that intact core particles do not enter the nucleus. To examine the role of the nucleus in core particle formation, we expressed p21.5 in surgically anucleate oocytes. We show that anucleate oocytes support efficient core particle formation, indicating that (i) the nucleus is not essential for assembly and (ii) the cytoplasm can assemble most core particles found in oocytes. On the basis of our data, we propose that in oocytes, most core particle assembly (up to 95%) occurs in the cytoplasm, but that at least approximately 5% of the cellular core particles are assembled in the nucleus and remain there. We discuss the implications of these findings for the formation of replication-competent core particles in infected cells.
Zhou
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SL
Hormone Research Institute, University of California, San Francisco 94143-0534.
Standring
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DN
D000595:Amino Acid Sequence; D000818:Animals; D002213:Capsid / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification; D002467:Cell Nucleus / Q000502:physiology; D003593:Cytoplasm / Q000502:physiology; D006515:Hepatitis B virus / Q000235:genetics*; D008845:Microinjections; D008969:Molecular Sequence Data; D009865:Oocytes / Q000502:physiology*; D012333:RNA, Messenger / Q000008:administration & dosage / Q000235:genetics*; D013462:Sulfur Radioisotopes; D014758:Viral Core Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification; D014981:Xenopus
D012333:RNA, Messenger; D013462:Sulfur Radioisotopes; D014758:Viral Core Proteins