Crystallization and X-ray diffraction of virus-like particles from a piscine betanodavirus.

ABSTRACT: Dragon grouper nervous necrosis virus (DGNNV), a member of the genus Betanodavirus, causes high mortality of larvae and juveniles of the grouper fish Epinephelus lanceolatus. Currently, there is no reported crystal structure of a fish nodavirus. The DGNNV virion capsid is derived from a single open reading frame that encodes a 338-amino-acid protein of approximately 37?kDa. The capsid protein of DGNNV was expressed to form virus-like particles (VLPs) in Escherichia coli. The VLP shape is T = 3 quasi-symmetric with a diameter of ?38?nm in cryo-electron microscopy images and is highly similar to the native virion. In this report, crystals of DGNNV VLPs were grown to a size of 0.27?mm within two weeks by the hanging-drop vapour-diffusion method at 283?K and diffracted X-rays to ?7.5?Å resolution. In-house X-ray diffraction data of the DGNNV VLP crystals showed that the crystals belonged to space group R32, with unit-cell parameters a = b = 353.00, c = 800.40?Å, ? = ? = 90, ? = 120°. 23?268 unique reflections were acquired with an overall Rmerge of 18.2% and a completeness of 93.2%. Self-rotation function maps confirmed the fivefold, threefold and twofold symmetries of the icosahedron of DGNNV VLPs.

SUBMITTER: Luo YC 

PROVIDER: S-EPMC4118809 | biostudies-literature | 2014 Aug

REPOSITORIES: biostudies-literature