Major and minor capsid proteins of human polyomavirus JC cooperatively accumulate to nuclear domain 10 for assembly into virions.
The human polyomavirus JC (JCV) replicates in the nuclei of infected cells. Here we report that JCV virions are efficiently assembled at nuclear domain 10 (ND10), which is also known as promyelocytic leukemia (PML) nuclear bodies. The major capsid protein VP1, the minor capsid proteins VP2 and VP3, and a regulatory protein called agnoprotein were coexpressed from a polycistronic expression vector in COS-7 cells. We found that VP1 accumulated to distinct subnuclear domains in the presence of VP2/VP3 and agnoprotein, while VP1 expressed alone was distributed both in the cytoplasm and in the nucleus. Mutation analysis revealed that discrete intranuclear accumulation of VP1 requires the presence of either VP2 or VP3. However, VP2 or VP3 expressed in the absence of VP1 showed diffuse, not discrete, nuclear localization. The C-terminal sequence of VP2/VP3 contains two basic regions, GPNKKKRRK (cluster 1) and KRRSRSSRS (cluster 2). The deletion of cluster 2 abolished the accumulation of VP1 to distinct subnuclear domains. Deletion of the C-terminal 34 residues of VP2/VP3, including both cluster 1 and cluster 2, caused VP1 to localize both in the cytoplasm and in the nucleus. Using immunoelectron microscopy of cells that coexpressed VP1, VP2/VP3, and agnoprotein, we detected the assembly of virus-like particles in discrete locations along the inner nuclear periphery. Both in oligodendrocytes of the human brain and in transfected cells, discrete nuclear domains for VP1 accumulation were identified as ND10, which contains the PML protein. These results indicate that major and minor capsid proteins cooperatively accumulate in ND10, where they are efficiently assembled into virions.
Shishido-Hara
Yukiko
Y
Department of Microbiology, Tokyo Metropolitan Institute for Neuroscience, Japan. shishido-hara@par.odn.ne.jp
Ichinose
Shizuko
S
Higuchi
Kayoko
K
Hara
Yoshinobu
Y
Yasui
Kotaro
K
D021581:Active Transport, Cell Nucleus; D000595:Amino Acid Sequence; D000818:Animals; D019556:COS Cells; D036022:Capsid Proteins / Q000235:genetics / Q000378:metabolism*; D002467:Cell Nucleus / Q000378:metabolism / Q000648:ultrastructure / Q000821:virology; D006801:Humans; D007577:JC Virus / Q000235:genetics / Q000502:physiology*; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D014162:Transfection; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism; D054334:Viral Regulatory and Accessory Proteins; D019065:Virus Assembly / Q000502:physiology*; D000086082:Viroporin Proteins
D036022:Capsid Proteins; D014764:Viral Proteins; D054334:Viral Regulatory and Accessory Proteins; C062481:VP1 protein, polyomavirus; C442543:VP2 protein, Polyomavirus; C042559:agnoprotein, polyomavirus; D000086082:Viroporin Proteins