Hepatitis C virus entry receptor dynamics and studies of platelet actin dynamics using lifeact-gfp: a potential role for actin nodules in filopodia formation

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http://eprints.org/relation/isVersionOf http://etheses.bham.ac.uk/id/document/16459 http://eprints.org/relation/isVolatileVersionOf http://etheses.bham.ac.uk/id/document/16459 http://eprints.org/relation/ispreviewThumbnailVersionOf http://etheses.bham.ac.uk/id/document/16459 16462 2 28280 document 16462 small.png image/png cd48265a210e2a8b3b5574a12f423caf MD5 7782 2011-11-23 13:18:58 http://etheses.bham.ac.uk//id/eprint/2995/5/small.png 2995 5 5 image/png thumbnail_small Thumbnail Documents conversion from application/pdf to thumbnail_small en public

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http://eprints.org/relation/isVersionOf http://etheses.bham.ac.uk/id/document/16458 http://eprints.org/relation/isVolatileVersionOf http://etheses.bham.ac.uk/id/document/16458 http://eprints.org/relation/isIndexCodesVersionOf http://etheses.bham.ac.uk/id/document/16458 archive 4031 disk0/00/00/29/95 2011-11-24 14:38:57 2019-01-23 09:55:17 2011-11-24 14:38:57 thesis show 0 Barnes Amy McKeating Jane A. Harris Helen University of Birmingham Hepatitis C virus entry receptor dynamics and studies of platelet actin dynamics using lifeact-gfp: a potential role for actin nodules in filopodia formation unpub RC 10col_mede Virus entry into target cells is mediated by the attachment of viral glycoproteins to cell surface receptors. Entry of the hepatitis C virus (HCV) into hepatocytes has been shown to require viral glycoprotein dimer E1E2 and the target cell receptor CD81, scavenger receptor-BI (SR-BI) and the tight junction proteins claudin-1 and occludin in an undefined mechanism. In this study, we aim to define the dynamic movements of these proteins in the membrane of the polarising hepatocarcinoma cell line HepG2. We further examine the extent to which these kinetics are perturbed following addition of the soluble viral glycoproteins sE2 and sE1E2, and the small molecule inhibitor of SR-BI ITX5061. The results of this study show that the amount of mobile receptor and the speed at which it diffuses varies according to its location within the cell. CD81 and claudin-1 are expressed equally in the filopodia and plasma membrane, whereas SR-BI is expressed at lower levels in the filopodia compared to the plasma membrane. We show that addition of both sE2 and sE1E2 has varying affects on both the speed and mobility of CD81 and claudin-1 and that the majority of significant effects observed for claudin-1 are observed at areas of potential cell contact. Finally, we demonstrate that addition of ITX5061 affects the diffusion coefficient of CD81 and CLDN-1 and the amount of mobile SR-BI. Furthermore, the effects on SR-BI are limited to areas of cell contact or exploratory regions. In summary, we present data which we hope will further current knowledge of the activity of these receptors in relation to their role in HCV infection. 2011-12 2011-08 graduation eng This unpublished thesis/dissertation is copyright of the author and/or third parties. The intellectual property rights of the author or third parties in respect of this work are as defined by The Copyright Designs and Patents Act 1988 or as modified by any successor legislation. Any use made of information contained in this thesis/dissertation must be in accordance with that legislation and must be properly acknowledged. Further distribution or reproduction in any format is prohibited without the permission of the copyright holder. public 123 University of Birmingham School of Immunity and Infection m_rs na 2011-12 graduation