Analysis of caveolar membrane transport on virus infection

最近的研究报道，无包膜病毒感染使用的是空泡内吞系统。我们发现人类冠状病毒229E是一种囊膜病毒，是一种已知引起感冒的病毒，它首先与CD13结合，然后移动到质膜上的小窝，并通过小窝进入细胞。然而，其作用机制还没有明确的界定。在本研究中，为了探讨空泡膜转运在人类冠状病毒229E感染中的作用，我们进行了以下实验：1)病毒包膜抗体的构建我们已经获得了8个针对病毒包膜跨膜蛋白S蛋白的抗体克隆，但任何克隆都不能在形态上起作用。2)荧光偶联病毒的构建我们建立了几个稳定表达荧光蛋白标记的N蛋白或S蛋白的宿主细胞。3)病毒受体的时移分析我们用荧光时移系统分析了人冠状病毒229E的受体CD13的行为。在本研究中，我们发现(1)CD13分子以流动的方式在质膜上滑动并聚集形成簇，(2)一些簇结合成更大的簇，(3)在质膜上可能存在最喜欢的聚集点。

Recent studies have reported that a caveolar endocytosis system is used by the non-enveloped virus infection. We found that humancoronavirus-229E, which is an enveloped virus and is known to be a cause of common cold, first binds to CD13, then moves to caveolae on the plasma membrane, and enters cells cell through caveolae. However, its mechanism is not clearly defined. In the present study, to examine how caveolar membrane transport is involved in the infection by humancoronavirus-229E, we performed experiments as follows.1) Construction of antibodies to virus envelopeWe have raised eight clones of antibodies to S-protein, which is the transmembrane protein in the virus envelope, but any clones could not work morphologically.2) Construction of fluorochorme-conjugated virusWe established several host cells that stably express the fluorescence-protein-tagged N-protein or S-protein. Now, we try to make fluorochorme-conjugated virus.3) Time lapse analysis of virus receptorWe have analyzed the behavior of cross-linked CD13, which is a receptor for humancoronavirus-229E, by fluorescence time-lapse system. In this study, we have found (1) that CD13 molecules slide on the plasma membrane as flow and accumulate to form clusters, (2) that some clusters are combined to become a larger cluster, and (3) that the favorite sites of clustering may exist in the plasma membrane.