Analysis of caveolar membrane transport on virus infection

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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.