Preliminary analysis of crystals of panicum mosaic virus (PMV) by X-ray diffraction and atomic force microscopy

Panicum mosaic virus (PMV), a spherical virus of diameter about 300 Å, has been crystallized in a form suitable for high-resolution structural analysis. The crystals were grown from 15% PEG 400 at room temperature and could be flash-frozen directly from their mother liquor. The crystals diffracted to beyond 2.7 Å resolution. A data set was collected at 100 K to an effective resolution of 3.2 Å [Weiss (2001), J. Appl. Cryst. 34, 130–135]. The crystals belonged to space group P2₁, with unit-cell parameters a = 411.7, b = 403.9, c = 412.5 Å, β = 89.7°. Self-rotation functions and molecular replacement with tobacco necrosis virus as the probe model yielded tentative positions and orientations for the two entire virus particles comprising the asymmetric unit and implied a pseudo-face-centered cubic packing arrangement. Investigation of lightly glutaraldehyde-fixed crystals in water using atomic force microscopy confirms the packing arrangement given by the molecular-replacement result. The images also show that contaminating virions of the satellite virus to PMV, known as satellite panicum mosaic virus (SPMV), can be incorporated into the PMV crystals by insertion into the interstices between PMV virions in the lattice. This is the first observation of such a phenomenon in macromolecular crystals.