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http://eprints.org/relation/isVolatileVersionOf http://etheses.bham.ac.uk/id/document/1653 http://eprints.org/relation/ispreviewThumbnailVersionOf http://etheses.bham.ac.uk/id/document/1653 http://eprints.org/relation/isVersionOf http://etheses.bham.ac.uk/id/document/1653 archive 4031 disk0/00/00/06/81 2010-05-19 11:06:49 2019-01-23 09:52:38 2011-07-19 16:21:27 thesis show 0 Matthews Sophie Anne Pallen Mark J. Bingle Lewis University of Birmingham unpub R1 QR180 QR 10col_mede Enterohaemorrhagic Escherichia coli (EHEC) O157:H7 is a formidable pathogen that uses a type-III secretion system to inject bacterial ‘effector’ proteins directly into host cells. Most effectors that are encoded within the locus of enterocyte effacement (LEE) have been studied extensively. This study aimed to characterise a selection of recently discovered non-LEE-encoded effectors using a variety of model systems. Firstly, a β-lactamase translocation assay was used to demonstrate translocation of novel effectors into host cells. The localisation of selected effectors was then investigated using mammalian cells and a yeast cell model. The effector EspM2 was shown to induce the formation of actin stress fibres in transfected HeLa cells and caused growth retardation when expressed in yeast. A number of NleG effectors also caused growth retardation and morphological changes when expressed in yeast. Growth retardation caused by the effector NleG8-2 was shown to be dependent on three conserved cysteine, aspartic acid and histidine residues. Transcriptomics and a high copy yeast gene suppression screen revealed that NleG8-2 may disrupt yeast physiology by affecting the secretory pathway. This study confirms that the effector repertoire of EHEC O157:H7 is much larger than previously imagined and provides insight into the function of selected novel effectors. 2010-07 2009-09 graduation eng author unchecked This unpublished thesis/dissertation is copyright of the author and/or third parties.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 279 University of Birmingham School of Immunity and Infection d_ph FALSE mrc 2010-07 graduation