Function of rafts and caveolae as virus infection

筏和小窝作为病毒感染的功能

• 批准号: 15590177
• 负责人: NOMURA Ryuji
• 金额: $ 2.3万
• 依托单位: Fujita Health University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2005
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15590177/
• 关键词: coronavirus; raft; caveola; cholesterol; caveolin; fibroblast

CD13, a receptor for human coronavirus (HCoV)-229E, was identified as a major component of the Triton X-100-resistant membrane microdomain in human fibroblasts. Incubation of living fibroblasts with an anti-CD13 antibody on ice gave punctate labeling that was evenly distributed on the cell surface, but raising the temperature to 37℃ before fixation caused aggregation of the labeling. The aggregated labeling of CD13 colocalized with caveolin-1 in most cells. The HCoV-229E virus particle showed a binding and redistribution pattern similar to the anti-CD13 antibody : the virus bound to the cell evenly when incubated on ice, but became colocalized with caveolin-1 at 37℃ ; importantly, the virus also caused sequestration of CD13 to the caveolin-1-positive area. Electron microscopy confirmed that HCoV-229E was localized near or at the orifice of caveolae after the incubation at 37℃. Depletion of plasmalemmal cholesterol with methyl □-cyclodextrin significantly reduced the HCoV-229E redistribution and the subsequent infection. Caveolin-1 knockdown by RNA interference also reduced the HCoV-229E infection considerably. The result indicates that HCoV-229E first binds to CD13 in the Triton X-100-resistant microdomain, clusters CD13 by cross-linking, and thereby reaches the caveolar region before entering cells.

CD13是人冠状病毒(HCoV)-229E的受体，被鉴定为人成纤维细胞Triton x -100抗性膜微域的主要成分。用抗cd13抗体在冰上孵育活成纤维细胞，可获得均匀分布在细胞表面的点状标记，但固定前将温度升高至37℃会导致标记聚集。在大多数细胞中，CD13的聚集标记与小窝蛋白-1共定位。HCoV-229E病毒颗粒表现出与抗cd13抗体相似的结合和再分布模式：在冰上孵卵时，病毒与细胞均匀结合，但在37℃时与小泡蛋白-1共定位；重要的是，该病毒还导致CD13被隔离到小窝蛋白-1阳性区域。电镜观察证实，37℃孵育后，HCoV-229E定位于小泡孔附近或孔处。甲基□-环糊精减少血浆胆固醇可显著降低HCoV-229E再分布和随后的感染。RNA干扰敲低Caveolin-1也能显著降低HCoV-229E感染。结果表明，HCoV-229E首先与Triton x -100抗性微域的CD13结合，通过交联聚集CD13，从而到达空泡区，然后进入细胞。

项目成果

Takao Senda: "The expression of cytokeratin in hepatic stellate cells of the cod."Arch.Histol.Cytol.. 66・5. 437-444 (2003)

Takao Senda：“鳕鱼肝星状细胞中细胞角蛋白的表达”。Arch.Histol.Cytol.. 66・5（2003）。

組織細胞化学2003

组织细胞化学2003

• 发表时间: 2003
• 作者: Takao Senda;et al.;日本組織細胞化学会
• 通讯作者: 日本組織細胞化学会

Anti-calreticulin Antibody Binds to a Membrane Protein in Caveolae

• DOI: 10.1267/ahc.38.43
• 发表时间: 2005
• 期刊: Acta Histochemica Et Cytochemica
• 影响因子: 2.4
• 作者: R. Nomura;T. Aoki;H. Hagiwara;T. Senda;T. Fujimoto

Human coronavirus 229E binds to CD13 in rafts and enters the cell through caveolae

• DOI: 10.1128/jvi.78.16.8701-8708.2004
• 发表时间: 2004-08-01
• 期刊: JOURNAL OF VIROLOGY
• 影响因子: 5.4
• 作者: Nomura, R;Kiyota, A;Fujimoto, T
• 通讯作者: Fujimoto, T

Lithium inhibits apoptosis of mouse neural progenitor cells

• DOI: 10.1097/00001756-200310060-00004
• 发表时间: 2003-10-06
• 期刊: NEUROREPORT
• 影响因子: 1.7
• 作者: Shimomura, A;Nomura, R;Senda, T
• 通讯作者: Senda, T