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水稻矮缩病毒非结构蛋白Pns11的核酸结合活性分析 被引量:1
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作者 程明非 任刚 +2 位作者 徐洪 陈章良 李毅 《高技术通讯》 EI CAS CSCD 2000年第9期1-4,共4页
水稻矮缩病毒的非结构蛋白Pns11是一个序列非特异性的核酸结合蛋白 ,其N端有类似锌指蛋白的结构 ,C端富含碱性氨基酸。为了研究Pns11的核酸结合活性 ,构建了两个位于可能的锌指结构内的点突变以及四个分别删除部分碱性区氨基酸片段的缺... 水稻矮缩病毒的非结构蛋白Pns11是一个序列非特异性的核酸结合蛋白 ,其N端有类似锌指蛋白的结构 ,C端富含碱性氨基酸。为了研究Pns11的核酸结合活性 ,构建了两个位于可能的锌指结构内的点突变以及四个分别删除部分碱性区氨基酸片段的缺失突变。经测序验证后分别克隆到大肠杆菌表达载体 pBV2 2 1中 ,温度诱导表达。用提取包涵体的方法初步纯化突变体蛋白 ,Western印迹检测这些突变体和Pns11的抗体均有特异反应。以Southwestern和Northwestern印迹的方法证明六个突变体都保持了核酸结合活性。结果表明N端的类似锌指蛋白的结构不是结合核酸的活性位点 ,部分缺失C端的碱性氨基酸也不影响其结合。 展开更多
关键词 水稻矮缩病毒 非结构蛋白pns11 点突变 缺失突变 核酸结合蛋白 活性
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A non-structural protein encoded by Rice Dwarf Virus targets to the nucleus and chloroplast and inhibits local RNA silencing
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作者 Feng Gao Shanshan Zhao +5 位作者 Shuzhen Men Zhensheng Kang Jian Hong Chunhong Wei Wei Hong Yi Li 《Science China(Life Sciences)》 SCIE CAS CSCD 2020年第11期1703-1713,共11页
RNA silencing is a potent antiviral mechanism in plants and animals. As a counter-defense, many viruses studied to date encode one or more viral suppressors of RNA silencing(VSR). In the latter case, how different VSR... RNA silencing is a potent antiviral mechanism in plants and animals. As a counter-defense, many viruses studied to date encode one or more viral suppressors of RNA silencing(VSR). In the latter case, how different VSRs encoded by a virus function in silencing remains to be fully understood. We previously showed that the nonstructural protein Pns10 of a Phytoreovirus, Rice dwarf virus(RDV), functions as a VSR. Here we present evidence that another nonstructural protein, Pns11, also functions as a VSR. While Pns10 was localized in the cytoplasm, Pns11 was localized both in the nucleus and chloroplasts. Pns11 has two bipartite nuclear localization signals(NLSs), which were required for nuclear as well as chloroplastic localization. The NLSs were also required for the silencing activities of Pns11. This is the first report that multiple VSRs encoded by a virus are localized in different subcellular compartments, and that a viral protein can be targeted to both the nucleus and chloroplast. These findings may have broad significance in studying the subcellular targeting of VSRs and other viral proteins in viral-host interactions. 展开更多
关键词 Rice dwarf virus pns11 RNA silencing suppressor nuclear and chloroplastic localization
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