Biochemical and physical characterization of parvovirus minute virus of mice virus-like particles.
The VP-2 major capsid protein of the prototype strain of the parvovirus minute virus of mice (MVMp) was expressed, using a baculovirus vector, in Sf9 insect cells. Immunogold electron microscopy of infected Sf9 cells showed VP-2 localized in the nucleus and cytoplasm as is observed in mammalian cells during natural infections. The VP-2 subunits self-assembled into empty parvovirus-like particles (VLPs), which appeared morphologically similar to and immunogenically indistinguishable from native empty MVMp particles, which also contain the minor capsid protein, VP1. Incubations under different pH and temperature conditions showed that the MVMp VLPs and native empty MVMp capsids share comparable stability. Once heated the particles can be similarly and specifically cleaved by trypsin at the VP-2 N-terminal domain. This process mimics the further maturation of the "rat-like" parvovirus virions, following viral DNA encapsidation, indicating that biologically relevant features of the MVMp capsid are maintained in the VLPs. Crystals have been obtained for the MVMp VLPs which were isomorphous to those used for the high-resolution structure determination of virions and native empty particles of the immunosuppressive strain of MVM (MVMi). The VLP crystals diffracted X rays to beyond 3-A resolution and are in space group C2 (a = 448.7, b = 416.6, c = 306.1 A, and beta = 95.9 degrees ). This is the first report of crystals from parvoviral particles produced in a heterologous system diffracting X rays to high resolution, indicating that VP-2 of some parvovirus capsids can self-assemble into ordered T = 1 icosahedral capsids in the absence of other viral and host cell functions.
Hernando
E
E
Centro de Biologia Molecular "Severo Ochoa" (UAM-CSIC), Universidad Autónoma de Madrid, Cantoblanco, Madrid, 28049, Spain.
Llamas-Saiz
A L
AL
Foces-Foces
C
C
McKenna
R
R
Portman
I
I
Agbandje-McKenna
M
M
Almendral
J M
JM
D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D002213:Capsid / Q000737:chemistry* / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D036022:Capsid Proteins; D002460:Cell Line; D003460:Crystallization; D005822:Genetic Vectors; D006358:Hot Temperature; D051379:Mice; D008854:Microscopy, Electron; D008915:Minute Virus of Mice / Q000737:chemistry* / Q000235:genetics / Q000648:ultrastructure; D018411:Spodoptera / Q000166:cytology / Q000235:genetics; D014771:Virion / Q000276:immunology / Q000378:metabolism / Q000648:ultrastructure; D014961:X-Ray Diffraction
D036022:Capsid Proteins; C442549:VP1 protein, Mice minute virus