Rice black-streaked dwarf virus outer capsid protein P10 has self-interactions and forms oligomeric complexes in solution
Rice black-streaked dwarf virus outer capsid protein P10 has self-interactions and forms oligomeric complexes in solution
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水稻黑条矮缩病毒外衣壳蛋白P10在溶液中存在自相互作用并形成寡聚复合物
DOI：
10.1016/j.virusres.2007.03.017
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发表时间：
2007-07-01
期刊：
影响因子：
5
通讯作者：
Li, Yi
中科院分区：
文献类型：
--
作者：
Liu, Huijun;Wei, Chunhong;Li, Yi
水稻黑条矮缩病毒(RBSDV)的S10 ORF编码的P10蛋白被认为是病毒颗粒外壳的组成部分。在本研究中，基于 GAL4 转录激活剂的酵母双杂交检测系统显示，P10 具有自我相互作用的能力，并通过体外远西方印迹分析进一步证实。负责 P10-P10 自相互作用的结构域被映射到蛋白质 N 末端区域的前 230 个氨基酸。使用杆状病毒表达系统中表达的纯化重组 P10 蛋白和戊二醛进行化学交联，进一步研究了 PI 0 的寡聚特性。在不存在其他病毒蛋白的情况下，完整的 P10 重组体主要以三聚体形式存在于溶液中，并表现出呼肠孤病毒科所有已知第二层蛋白单元所共有的寡聚性质。编码前 230 个 N 端氨基酸的截短 PI 0 突变体失去了形成三聚体的能力，尽管在交联测定过程中检测到二聚体形式。还原或非还原条件下的聚丙烯酰胺凝胶电泳表明，P10 亚基不是通过分子间二硫键寡聚化，而是可能通过一些其他类型的缔合，例如疏水或电荷相互作用。 (C) 2007 Elsevier B.V. 保留所有权利。
The P10 protein encoded by S10 ORF of Rice black-streaked dwarf virus (RBSDV) was thought to be the component of outer shell of viral particle. In the present study, P10 has an ability for self-interaction as shown by a GAL4 transcription activator-based yeast two-hybrid assay system and further confirmed by in vitro far-Western blot analysis. The domain responsible for P10-P10 self-interaction was mapped to the first 230 amino acids at the N-terminal region of the protein. The oligomerization property of PI 0 was further investigated using chemical cross-linking with purified recombinant P10 proteins expressed in a baculovirus expression system and glutaraldehyde. Intact P10 recombinants existed predominantly as trimers in solution in the absence of other viral proteins and displayed the oligomeric nature common to all known second-layer protein units of the Reoviridae. A truncated PI 0 mutant encoding the first 230 N-terminal amino acids lost its ability to form trimers even though dimeric, forms were detected during the cross-linking assay. Polyacrylamide gel electrophoresis under reducing or non-reducing conditions suggested that P10 subunits were oligomerized not through intermolecular disulfide bonds, but perhaps through some other type of association, such as hydrophobic or charge interactions. (C) 2007 Elsevier B.V. All rights reserved.