首页> 外文期刊>Acta crystallographica, Section F. Structural biology and crystallization communications >Crystallization and preliminary crystallographic analysis of a viral RNA-silencing suppressor encoded by Wuhan nodavirus
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Crystallization and preliminary crystallographic analysis of a viral RNA-silencing suppressor encoded by Wuhan nodavirus

机译:武汉诺达病毒编码的病毒RNA沉默抑制剂的结晶和初步晶体学分析

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摘要

Wuhan nodavirus (WhNV), which is a new member of the Nodaviridae family, encodes a viral protein, B2, that suppresses RNA silencing and host-cell RNA interference (RNAi)-mediated immunity. Although Flock House virus (FHV), another member of the Nodaviridae family, also produces a B2 protein with a similar function, the primary sequences of the B2 proteins from WhNV and FHV have no similarity. To gain a better understanding of the structural details and the mechanism of suppression of RNA silencing by WhNV B2 and to compare it with FHV B2, recombinant WhNV B2 protein has been overexpressed in Escherichia coli, purified and crystallized at 291 K using PEG 4000 as a precipitant. A 2.8 angstrom resolution data set has been collected from a single crystal at 100 K. This crystal belonged to space group P2(1)2(1)2(1), with unit-cell parameters a = 27.3, b = 45.6, c = 133.9 angstrom, alpha = beta = gamma = 90 degrees. Assuming the presence of two molecules in the asymmetric unit, the Matthews coefficient is 2.2 angstrom(3) Da(-1).
机译:武汉诺达病毒(WhNV)是诺达病毒科的新成员,它编码一种病毒蛋白B2,该蛋白可抑制RNA沉默和宿主细胞RNA干扰(RNAi)介导的免疫。尽管Nodaviridae家族的另一成员Flock House病毒(FHV)也产生具有相似功能的B2蛋白,但来自WhNV和FHV的B2蛋白的主要序列却没有相似性。为了更好地理解WhNV B2抑制RNA沉默的结构细节和机制并将其与FHV B2进行比较,重组WhNV B2蛋白已在大肠杆菌中过表达,并在291 K下使用PEG 4000作为纯化和结晶。沉淀剂。从100 K的单晶收集了2.8埃分辨率数据集。该晶体属于空间群P2(1)2(1)2(1),单位像元参数a = 27.3,b = 45.6,c = 133.9埃,α=β=伽马= 90度。假设在不对称单元中存在两个分子,则马修斯系数为2.2埃(3)Da(-1)。

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