SARS coronavirus unique domain: three-domain molecular architecture in solution and RNA binding.
SARS coronavirus unique domain: three-domain molecular architecture in solution and RNA binding.
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DOI：
10.1016/j.jmb.2010.05.027
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发表时间：
2010-07-23
影响因子：
5.6
通讯作者：
Wüthrich K
中科院分区：
文献类型：
--
作者：
Johnson MA;Chatterjee A;Neuman BW;Wüthrich K
严重急性呼吸综合征（SARS）冠状病毒的非结构蛋白3包括一个“SARS独特结构域”（SUD），由三个由短连接肽段分隔的球形结构域组成。本文报道了c端结构域（SUD-C）和包含中间结构域（SUD-M）和c端结构域的双结构域（SUD-MC）的NMR结构测定，以及n端结构域（SUD-N）和SUD-NM双结构域的构象态的NMR数据。在溶液中，SUD-N和SUD-NM均为单体和球状；在SUD-NM中，两个残基间结构域连接序列具有高迁移率，两个结构域没有优先的相对取向。SUD-C采用类似于dna修饰酶的dna结合结构域，结构上与dna修饰酶相似。SUD-M（先前确定的）和SUD-C（来自本研究的）的结构在SUD-MC中保持不变，其中两个结构域是灵活连接的。凝胶移位实验表明，与嘧啶碱基相比，SUD-C和SUD-MC与单链RNA的结合和对嘌呤碱基的识别更强，因此SUD-MC与含有嘌呤的RNA序列的结合比SUD-M更受限制。通过对15n标记蛋白的观察，核磁共振化学位移微扰实验进一步得出了RNA结合位点的描述（即，在SUD-M中，一个带明显空腔的带正电的表面区域，在SUD-C中，一个反平行β片的几个残基）。总的来说，目前的数据为涉及SUD- m和SUD- c协同作用的分子机制提供了证据，这些机制导致可能是SUD和SARS冠状病毒所特有的特异性RNA结合。
Nonstructural protein 3 of the severe acute respiratory syndrome (SARS) coronavirus includes a “SARS-unique domain” (SUD) consisting of three globular domains separated by short linker peptide segments. This work reports NMR structure determinations of the C-terminal domain (SUD-C) and a two-domain construct (SUD-MC) containing the middle domain (SUD-M) and the C-terminal domain, and NMR data on the conformational states of the N-terminal domain (SUD-N) and the SUD-NM two-domain construct. Both SUD-N and SUD-NM are monomeric and globular in solution; in SUD-NM, there is high mobility in the two-residue interdomain linking sequence, with no preferred relative orientation of the two domains. SUD-C adopts a frataxin like fold and has structural similarity to DNA-binding domains of DNA-modifying enzymes. The structures of both SUD-M (previously determined) and SUD-C (from the present study) are maintained in SUD-MC, where the two domains are flexibly linked. Gel-shift experiments showed that both SUD-C and SUD-MC bind to single-stranded RNA and recognize purine bases more strongly than pyrimidine bases, whereby SUD-MC binds to a more restricted set of purine-containing RNA sequences than SUD-M. NMR chemical shift perturbation experiments with observations of 15N-labeled proteins further resulted in delineation of RNA binding sites (i.e., in SUD-M, a positively charged surface area with a pronounced cavity, and in SUD-C, several residues of an anti-parallel β-sheet). Overall, the present data provide evidence for molecular mechanisms involving the concerted actions of SUD-M and SUD-C, which result in specific RNA binding that might be unique to the SUD and, thus, to the SARS coronavirus.
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影响因子：
5
作者：
Gorbalenya AE;Enjuanes L;Ziebuhr J;Snijder EJ
通讯作者：
Snijder EJ
影响因子：
5.7
作者：
Nair, M;Adinolfi, S;Pastore, A
通讯作者：
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影响因子：
64.8
作者：
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通讯作者：
Tang, WJ
影响因子：
--
作者：
Masters PS
通讯作者：
Masters PS
DOI：
10.1073/pnas.0504818102
发表时间：
2005-08-02
影响因子：
11.1
作者：
Hiller, S;Fiorito, F;Wider, G
通讯作者：
Wider, G