Expression, crystallization and preliminary crystallographic analysis of RNA-binding protein Hfq (YmaH) from Bacillus subtilis in complex with an RNA aptamer
Expression, crystallization and preliminary crystallographic analysis of RNA-binding protein Hfq (YmaH) from Bacillus subtilis in complex with an RNA aptamer
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DOI：
10.1107/s1744309110009942
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发表时间：
2010-05-01
影响因子：
0.9
通讯作者：
Kumasaka, Takashi
中科院分区：
文献类型：
--
作者：
Baba, Seiki;Someya, Tatsuhiko;Kumasaka, Takashi
Hfq蛋白是一种六聚体RNA结合蛋白，在不同的环境胁迫下通过与RNA结合来调节基因的表达。它的环状结构结合了各种类型的RNA，包括mRNA和sRNA。大肠杆菌和金黄色葡萄球菌Hfq的RNA结合结构分别在远端含有Poly(A)RNA，在近端含有富含U的RNA。本文报道了利用悬滴蒸气扩散技术制备的枯草芽孢杆菌Hfq蛋白与A/G-重复7聚体RNA(Hfq-RNA)的复合体的晶体。1型Hfq-RNA晶体属于I422空间群，晶胞参数a=b=123.70，c=119.13埃；2型Hfq-RNA晶体属于F222空间群，晶胞参数a=91.92，b=92.50，c=114.92埃。从两种晶型收集了2.20埃的分辨率的衍射数据。自旋分析清楚地显示了Hfq蛋白的六聚体结构。
The Hfq protein is a hexameric RNA-binding protein which regulates gene expression by binding to RNA under the influence of diverse environmental stresses. Its ring structure binds various types of RNA, including mRNA and sRNA. RNA-bound structures of Hfq from Escherichia coli and Staphylococcus aureus have been revealed to have poly(A) RNA at the distal site and U-rich RNA at the proximal site, respectively. Here, crystals of a complex of the Bacillus subtilis Hfq protein with an A/G-repeat 7-mer RNA (Hfq-RNA) that were prepared using the hanging-drop vapour-diffusion technique are reported. The type 1 Hfq-RNA crystals belonged to space group I422, with unit-cell parameters a = b = 123.70, c = 119.13 angstrom, while the type 2 Hfq-RNA crystals belonged to space group F222, with unit-cell parameters a = 91.92, b = 92.50, c = 114.92 angstrom. Diffraction data were collected to a resolution of 2.20 angstrom from both crystal forms. The hexameric structure of the Hfq protein was clearly shown by self-rotation analysis.