Preliminary analysis of two and three dimensional crystals of vault ribonucleoprotein particles.
Vaults are large ribonucleoprotein particles found in a wide variety of eukaryotes. When imaged by electron-microscopy vaults present a strikingly conserved barrel-shaped structure with an invaginated waist and two protruding caps. In this work, we present two dimensional (2D) and three dimensional (3D) crystals of naturally produced vaults in murine and monkey cells, respectively. The 2D-crystals presented a hexagonal packing with the lattice parameter defined by the diameter of the vault barrel. Fourier transforms from images of the negatively stained 2D-crystals showed spots till about 45 A resolution. The 3D-crystals reached about 0.15 x 0.15 x 0.02 mm3 in size and presented a flat triangular morphology with well-developed faces. The preliminary characterization of these 3D-crystals, which diffract very weakly to approximately 10 A resolution, suggests a trigonal packing with the R32 space group symmetry. The 3D-crystals appear to be formed by adding layers of vaults, which retain the hexagonal organization seen in the 2D-crystals, with relative shifts that maximize the interdigitation of particles in adjacent layers. Accurate crystal symmetry in the 2D- and 3D-crystals requires neighbor particles interacting according to a 6-fold and a 3-fold dihedral symmetry, respectively. Compatibility with the reported 8-fold symmetry would imply multiples of 24-fold rotational symmetry, in agreement with the recently proposed 48-fold dihedral symmetry for reconstituted recombinant vaults.
Querol-Audí
Jordi
J
Instituto de Biología Molecular de Barcelona (CSIC-Parc Científic), Josep-Samitier 1-5, Barcelona 08028, Spain.
Perez-Luque
Rosa
R
Fita
Ignacio
I
Lopéz-Iglesias
Carmen
C
Castón
José R
JR
Carrascosa
José L
JL
Verdaguer
Nuria
N
D000818:Animals; D002478:Cells, Cultured; D003460:Crystallization; D018360:Crystallography, X-Ray; D000882:Haplorhini; D051379:Mice; D008854:Microscopy, Electron; D020394:Vault Ribonucleoprotein Particles / Q000737:chemistry* / Q000648:ultrastructure*
D020394:Vault Ribonucleoprotein Particles