Crystallization and preliminary X-ray crystallographic analysis of ribosome assembly factors: the Rpf2-Rrs1 complex
Crystallization and preliminary X-ray crystallographic analysis of ribosome assembly factors: the Rpf2-Rrs1 complex
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DOI：
10.1107/s2053230x14024182
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发表时间：
2014-12-01
影响因子：
0.9
通讯作者：
Yao, Min
中科院分区：
文献类型：
--
作者：
Asano, Nozomi;Nakamura, Akiyoshi;Yao, Min
Rpf2和Rrs1是核糖体生物发生的必需蛋白。这些蛋白与5S rRNA和两个核糖体蛋白（L5和L11）形成复合体（rpf2亚复合体）。该复合体被招募到核糖体前体（90S核糖体前体）。这种招募对于25S rRNA的成熟是必要的。Rpf2和Rrs1的基因缺失导致25S rRNA前体的积累。本研究将来自细粒曲霉的Rpf2和Rrs1在大肠杆菌中共过表达，并进行纯化和结晶。随后的分析表明，这些晶体含有由两个n端结构域组成的复合物的中心核心区域。采集了分辨率为2.35埃的x射线衍射数据。初步分析表明，该晶体属于P2(1)2(1)2(1)空间群，晶胞参数a = 54.1, b = 123.3, c = 133.8埃。不对称单元中有两个配合物。硒代蛋氨酸标记蛋白的结构测定正在进行中。
Rpf2 and Rrs1 are essential proteins for ribosome biogenesis. These proteins form a complex (the Rpf2-subcomplex) with 5S rRNA and two ribosomal proteins (L5 and L11). This complex is recruited to the ribosome precursor (the 90S pre-ribosome). This recruitment is necessary for the maturation of 25S rRNA. Genetic depletion of Rpf2 and Rrs1 results in accumulation of the 25S rRNA precursor. In this study, Rpf2 and Rrs1 from Aspergillus nidulans were co-overexpressed in Escherichia coli, purified and crystallized. Subsequent analysis revealed that these crystals contained the central core region of the complex consisting of both N-terminal domains. X-ray diffraction data were collected to 2.35 angstrom resolution. Preliminary analysis revealed that the crystals belonged to space group P2(1)2(1)2(1), with unit-cell parameters a = 54.1, b = 123.3, c = 133.8 angstrom. There are two complexes in the asymmetric unit. Structure determination using selenomethionine-labelled protein is in progress.