title: 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, α = β = 90, γ = 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.
authors.affiliation: Department of Marine Biotechnology and Resources, National Sun Yat-sen University, Kaohsiung 80424, Taiwan.
authors.forename: Yu-Chun
authors.initials: YC
authors.lastname: Luo
authors.affiliation: Department of Marine Biotechnology and Resources, National Sun Yat-sen University, Kaohsiung 80424, Taiwan.
authors.forename: Chun-Hsiung
authors.initials: CH
authors.lastname: Wang
authors.affiliation: Department of Marine Biotechnology and Resources, National Sun Yat-sen University, Kaohsiung 80424, Taiwan.
authors.forename: Yi-Min
authors.initials: YM
authors.lastname: Wu
authors.affiliation: Department of Marine Biotechnology and Resources, National Sun Yat-sen University, Kaohsiung 80424, Taiwan.
authors.forename: Wangta
authors.initials: W
authors.lastname: Liu
authors.affiliation: Department of Marine Biotechnology and Resources, National Sun Yat-sen University, Kaohsiung 80424, Taiwan.
authors.forename: Ming-Wei
authors.initials: MW
authors.lastname: Lu
authors.affiliation: Department of Marine Biotechnology and Resources, National Sun Yat-sen University, Kaohsiung 80424, Taiwan.
authors.forename: Chan-Shing
authors.initials: CS
authors.lastname: Lin
mesh_terms: D003460:Crystallization; D008854:Microscopy, Electron; D030381:Nodaviridae / Q000737:chemistry*; D014771:Virion / Q000737:chemistry*; D014961:X-Ray Diffraction / Q000379:methods*
keywords: Betanodavirus; Dragon grouper nervous necrosis virus (DGNNV); Epinephelus lanceolatus
references.citation: Virology. 2001 Nov 10;290(1):50-8
references.pmid: 11883005
references.citation: J Mol Biol. 1987 Aug 5;196(3):591-7
references.pmid: 2824786
references.citation: Methods Enzymol. 1997;276:307-26
references.pmid: 27754618
references.citation: Methods. 2004 Nov;34(3):415-23
references.pmid: 15325658
references.citation: Acta Crystallogr D Biol Crystallogr. 2005 Aug;61(Pt 8):1041-8
references.pmid: 16041068
references.citation: J Gen Virol. 2003 Jun;84(Pt 6):1577-1582
references.pmid: 12771428
references.citation: Appl Environ Microbiol. 1997 Apr;63(4):1633-6
references.pmid: 9097459
references.citation: Nat Struct Biol. 2001 Jan;8(1):77-83
references.pmid: 11135676
references.citation: Virology. 1992 Mar;187(1):368-71
references.pmid: 1736540
references.citation: Virus Genes. 2010 Aug;41(1):73-80
references.pmid: 20446029
references.citation: Vaccine. 2006 Sep 11;24(37-39):6282-7
references.pmid: 16797803
references.citation: J Mol Biol. 1968 Apr 28;33(2):491-7
references.pmid: 5700707
references.citation: Arch Virol. 2008;153(9):1633-42
references.pmid: 18626568
references.citation: Proc Natl Acad Sci U S A. 1965 Aug;54(2):513-21
references.pmid: 5217439
references.citation: Acta Crystallogr B. 1992 Aug 1;48 ( Pt 4):515-20
references.pmid: 1418822
references.citation: Acta Crystallogr D Biol Crystallogr. 2005 Apr;61(Pt 4):490-3
references.pmid: 15805606
references.citation: J Virol. 2002 Jun;76(12):6370-5
references.pmid: 12021370
references.citation: Methods Enzymol. 2003;368:188-216
references.pmid: 14674275