Crystallization of Brome mosaic virus and T = 1 Brome mosaic virus particles following a structural transition.
Brome mosaic virus (BMV), a T = 3 icosahedral plant virus, can be dissociated into coat protein subunits and subunit oligomers at pH 7.5 in the presence of concentrated salts. We have found that during the course of this treatment the coat protein subunits are cleaved, presumably by plant cell proteases still present in the preparation, between amino acids 35 and 36. The truncated protein subunits will then reorganize into T = 1 icosahedral particles and can be crystallized from sodium malonate. Quasi elastic light scattering and atomic force microscopy results suggest that the transition from T = 3 to T = 1 particles can occur by separate pathways, dissociation into coat protein subunits and oligomers and reassembly into T = 1 particles, or direct condensation of the T = 3 virions to T = 1 particles with the shedding of hexameric capsomeres. The latter process has been directly visualized using atomic force microscopy. Native T = 3 virions have been crystallized in several different crystal forms, but neither a rhombohedral form nor either of two orthorhombic forms diffract beyond about 3.4 A. Tetragonal crystals of the T = 1 particles, however, diffract to at least 2.5 A resolution. Evidence suggests that the T = 1 particles are more structurally uniform and ordered than are native T = 3 virions. A variety of anomalous virus particles having diverse sizes have been visualized in preparations of BMV used for crystallization. In some cases these aberrant particles are incorporated into growing crystals where they are frequently responsible for defect formation.
Lucas
R W
RW
Department of Molecular Biology and Biochemistry, University of California, Irvine, 92697-3900, USA.
Kuznetsov
Y G
YG
Larson
S B
SB
McPherson
A
A
D017795:Bromovirus / Q000737:chemistry* / Q000378:metabolism / Q000648:ultrastructure*; D003460:Crystallization; D018360:Crystallography, X-Ray / Q000379:methods; D001467:Hordeum / Q000821:virology; D008027:Light; D018625:Microscopy, Atomic Force; D012542:Scattering, Radiation; D014771:Virion / Q000737:chemistry* / Q000378:metabolism / Q000648:ultrastructure*