Preliminary X-ray data analysis of crystalline hibiscus chlorotic ringspot virus.
Hibiscus chlorotic ringspot virus (HCRSV) is a positive-sense monopartite single-stranded RNA virus that belongs to the Carmovirus genus of the Tombusviridae family, which includes carnation mottle virus (CarMV). The HCRSV virion has a 30 nm diameter icosahedral capsid with T = 3 quasi-symmetry containing 180 copies of a 38 kDa coat protein (CP) and encapsidates a full-length 3.9 kb genomic RNA. Authentic virus was harvested from infected host kenaf leaves and was purified by saturated ammonium sulfate precipitation, sucrose density-gradient centrifugation and anion-exchange chromatography. Virus crystals were grown in multiple conditions; one of the crystals diffracted to 3.2 A resolution and allowed the collection of a partial data set. The crystal belonged to space group R32, with unit-cell parameters a = b = 336.4, c = 798.5 A. Packing considerations and rotation-function analysis determined that there were three particles per unit cell, all of which have the same orientation and fixed positions, and resulted in tenfold noncrystallography symmetry for real-space averaging. The crystals used for the structure determination of southern bean mosaic virus (SBMV) have nearly identical characteristics. Together, these findings will greatly aid the high-resolution structure determination of HCRSV.
Cheng
Ao
A
Department of Biological Sciences, National University of Singapore, 14 Science Drive 4, Kent Ridge, Singapore 117543, Singapore.
Speir
Jeffrey A
JA
Yuan
Y Adam
YA
Johnson
John E
JE
Wong
Sek Man
SM
D000595:Amino Acid Sequence; D002213:Capsid / Q000737:chemistry / Q000648:ultrastructure; D036022:Capsid Proteins / Q000737:chemistry / Q000648:ultrastructure; D019184:Carmovirus / Q000737:chemistry* / Q000302:isolation & purification / Q000648:ultrastructure; D003460:Crystallization; D003625:Data Collection; D019281:Dimerization; D031584:Hibiscus / Q000821:virology*; D008027:Light; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010316:Particle Size; D018515:Plant Leaves / Q000821:virology; D017510:Protein Folding; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D012367:RNA, Viral / Q000737:chemistry / Q000648:ultrastructure; D012399:Rotation; D012542:Scattering, Radiation; D017386:Sequence Homology, Amino Acid; D013223:Statistics as Topic; D013696:Temperature; D014771:Virion / Q000737:chemistry / Q000648:ultrastructure; D014961:X-Ray Diffraction
D036022:Capsid Proteins; D012367:RNA, Viral