Analysis of the nucleocapsid proteins that are constitutively expressed in cell lines
Analysis of the nucleocapsid proteins that are constitutively expressed in cell lines
批准号:
09670312
负责人:
NISHIO Machiko
金额:
$2.05万
依托单位:
依托单位国家:
日本
项目类别:
Grant-in-Aid for Scientific Research (C)
财政年份:
1997
资助国家:
日本
项目状态:
已结题
起止时间:
1997 至 1999
中文摘要
1.我已经分离了许多针对hPIV-2的P或NP蛋白的单克隆抗体。然而,尚未获得特异于V或L蛋白的MAbs。因此,本研究制备了抗V或抗L蛋白的单克隆抗体,并证明了V或L蛋白在病毒感染细胞中的分布.用抗P单克隆抗体和P蛋白缺失突变体进行Western免疫印迹,确定了P-P相互作用所必需的区域。P蛋白211-248位氨基酸区域是正确折叠和同源三聚体所必需的,而同源三聚体是由该区域的卷曲螺旋介导的.通过使用抗NP单克隆抗体的Western免疫印迹,对NP蛋白的缺失突变体进行分析,以鉴定NP-NP相互作用所必需的区域和NP蛋白上的P蛋白结合位点。结果表明,NP蛋白N端294个氨基酸都是NP-Np自组装所必需的,而P蛋白C端的两个氨基酸和另一个氨基酸是NP-Np自组装所必需的。aa 403 -494,连接到P蛋白的N端部分。通过使用抗L单克隆抗体的Western免疫印迹,我证明了L蛋白直接结合到P和NP蛋白。此外,确定了L蛋白结合P或NP蛋白的必需区域。P蛋白上的aa 278-353区域是与L蛋白结合所必需的。NP蛋白上的氨基酸403-494区域是与L蛋白结合所必需的.通过Northwestern印迹分析,我证明了P,V或NP蛋白上与RNA结合的区域。
英文摘要
1. I have already isolated a lot of monoclonal antibodies directed against the P or NP protein of hPIV-2. However, no MAbs specific for the V or L protein has been obtained. Therefore, I prepared anti-V or anti-L protein MAbs, and demonstrated the distribution of the V or L protein in virus-infected cells.2. By Western immunoblotting with anti-P MAbs and P protein deletion mutants, the region essential for P-P interactions was determined. The P protein region encompassing aa 211-248 was required for proper folding and homotrimer which is mediated by predicted coiled-coils in this region.3. By Western immunoblotting using anti-NP MAbs, the analysis of deletion mutants of the NP protein was performed to identify the region essential for NP-NP interaction and P protein-binding sites on the NP protein. The results indicate that the N-terminal 294 aa of the NP protein are all required for NP-Np self-assembly, and that two C-terminal parts of the P protein and another region. aa403-494, to the N-terminal part of the P protein.4. By Western immunoblotting using anti-L MAbs, I demonstrated that the L protein directly binds to the P and NP proteins. Furthermore, the essential region for the L protein binding on the P or NP protein was determined. The region aa 278-353 on the P protein is required for binding to the L protein. And the region aa 403-494 on the NP protein is required for binding to the L protein.5. By Northwestern blot assay, I demonstrated the regions on the P, V or NP protein which binds to RNA.
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Kousuke Okamoto: "Enhancement of himan parainfluenza virus-induced cell fusion by pradimicin, a low molecular weight mannose-binding antibiotic"Medical Microbiology and Immunology. Vol.186. 101-108 (1997)
Kousuke Okamoto:“pradimicin(一种低分子量甘露糖结合抗生素)增强希曼副流感病毒诱导的细胞融合”医学微生物学和免疫学。
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Morihiro Ito: "Role of a single amino acid at the amino terminus of the simian virus 5 F2 subunit in syncytium formation." Journal of Virology. Vol.71. 9855-9858 (1997)
Morihiro Ito:“猿病毒 5 F2 亚基氨基末端的单个氨基酸在合胞体形成中的作用。”
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Shigeru Suga: "Human immunodeficiency virus type-1 envelope glycoprotein gp120 induces expression of fusion regulatory protein(FRP)-1/CD98 on CD4+T cells." Medical Microbiology and Immunology. Vol.186. 237-243 (1997)
Shigeru Suga:“人类免疫缺陷病毒 1 型包膜糖蛋白 gp120 诱导 CD4 T 细胞上融合调节蛋白 (FRP)-1/CD98 的表达。”
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作者:
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通讯作者:
Morihiro Ito: "Role of a single amino acid at the amino terminus of the simian virus 5 F2 subunit syncytium formation"Journal of Virology. Vol.71. 9855-9858 (1997)
伊藤守宏:“猿猴病毒5 F2亚基合胞体形成的氨基末端单个氨基酸的作用”病毒学杂志。
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Machiko Nishio: "Mapping of domains on the human parainfluenza virus type 2 nucleocapsid protein (NP) required for NP-phosphoprotein or NP-NP interaction"Journal of General Virology. Vol.80. 2017-2022 (1999)
Machiko Nishio:“NP-磷蛋白或 NP-NP 相互作用所需的人副流感病毒 2 型核衣壳蛋白 (NP) 上的结构域图谱”普通病毒学杂志。
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共 11 条
Analysis of virus replication mechanism using minigenome or recombinant virus system
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批准号:23590539
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项目类别:Grant-in-Aid for Scientific Research (C)
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资助金额:$3.33万
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财政年份:2011
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负责人:NISHIO Machiko
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依托单位:
Comprehensive search and analysis of host factors that interact with the V protein of human parainfluenza virus type 2
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批准号:20590469
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项目类别:Grant-in-Aid for Scientific Research (C)
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资助金额:$3.0万
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财政年份:2008
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负责人:NISHIO Machiko
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依托单位:
Analysis of the function of paramyxovaus V protein
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批准号:18590448
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项目类别:Grant-in-Aid for Scientific Research (C)
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资助金额:$2.55万
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财政年份:2006
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负责人:NISHIO Machiko
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依托单位:
Analysis of the function of hPIV2 V protein by using reverse genetics.
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批准号:15590416
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项目类别:Grant-in-Aid for Scientific Research (C)
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资助金额:$2.3万
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财政年份:2003
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负责人:NISHIO Machiko
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依托单位:
海外基金