title: Role of the C-terminal region of vervet monkey polyomavirus 1 VP1 in virion formation.
abstract: Recently, we detected novel vervet monkey polyomavirus 1 (VmPyV) in a vervet monkey. Among amino acid sequences of major capsid protein VP1s of other polyomaviruses, VmPyV VP1 is the longest with additional amino acid residues in the C-terminal region. To examine the role of VmPyV VP1 in virion formation, we generated virus-like particles (VLPs) of VmPyV VP1, because VLP is a useful tool for the investigation of the morphological characters of polyomavirus virions. After the full-length VmPyV VP1 was subcloned into a mammalian expression plasmid, the plasmid was transfected into human embryonic kidney 293T (HEK293T) cells. Thereafter, VmPyV VLPs were purified from the cell lysates of the transfected cells via sucrose gradient sedimentation. Electron microscopic analyses revealed that VmPyV VP1 forms VLPs with a diameter of approximately 50 nm that are exclusively localized in cell nuclei. Furthermore, we generated VLPs consisting of the deletion mutant VmPyV VP1 (ΔC VP1) lacking the C-terminal 116 amino acid residues and compared its VLP formation efficiency and morphology to those of VLPs from wild-type VmPyV VP1 (WT VP1). WT and ΔC VP1 VLPs were similar in size, but the number of ΔC VP1 VLPs was much lower than that of WT VP1 VLPs in VP1-expressing HEK293T cells. These results suggest that the length of VP1 is unrelated to virion morphology; however, the C-terminal region of VmPyV VP1 affects the efficiency of its VLP formation.
authors.affiliation: Division of Molecular Pathobiology, Research Center for Zoonosis Control, Hokkaido University, Sapporo, Hokkaido 001-0020, Japan.
authors.forename: Hiroki
authors.initials: H
authors.lastname: Yamaguchi
authors.forename: Shintaro
authors.initials: S
authors.lastname: Kobayashi
authors.forename: Junki
authors.initials: J
authors.lastname: Maruyama
authors.forename: Michihito
authors.initials: M
authors.lastname: Sasaki
authors.forename: Ayato
authors.initials: A
authors.lastname: Takada
authors.forename: Takashi
authors.initials: T
authors.lastname: Kimura
authors.forename: Hirofumi
authors.initials: H
authors.lastname: Sawa
authors.forename: Yasuko
authors.initials: Y
authors.lastname: Orba
mesh_terms: D000818:Animals; D036022:Capsid Proteins / Q000235:genetics* / Q000648:ultrastructure; D002522:Chlorocebus aethiops / Q000821:virology*; D057809:HEK293 Cells; D006801:Humans; D015151:Immunoblotting; D007150:Immunohistochemistry; D008854:Microscopy, Electron; D010957:Plasmids / Q000235:genetics; D011120:Polyomavirus / Q000235:genetics*; D014162:Transfection; D014771:Virion / Q000235:genetics*
chemical_list: D036022:Capsid Proteins; C062481:VP1 protein, polyomavirus