Structural characterisation of the interaction between RBBP6 and the multifunctional protein YB-1

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dc.contributor.advisorPugh, David J.R.
dc.contributor.authorMuleya, Victor
dc.contributor.otherDept. of Biotechnology
dc.contributor.otherFaculty of Science
dc.date.accessioned2013-10-24T09:04:39Z
dc.date.accessioned2024-05-09T08:19:17Z
dc.date.available2011/05/13 08:05
dc.date.available2011/05/13
dc.date.available2013-10-24T09:04:39Z
dc.date.available2024-05-09T08:19:17Z
dc.date.issued2010
dc.descriptionMagister Scientiae - MScen_US
dc.description.abstractAs a means of further localising the interaction, truncated fragments derived from the C-terminal region of YB-1, were tested for their interaction with the RING finger domain of RBBP6 using three different assays: a directed yeast 2-hybrid assay, co-immunoprecipitation and NMR chemical shift perturbation analysis. Our results suggest that the entire 62 amino acid region at the C-terminal domain of YB-1 may be involved in the interaction with RBBP6. Using chemical shift perturbation analysis, this study provides an indication of where YB-1 binds to the RING finger. This represents the first step towards the design of therapeutics aimed at modulating the interaction between RBBP6 and YB-1 as a means of targeting the oncogenic effects of YB-1. In order to identify E2 enzymes involved in the ubiquitination of YB-1, we examined the efficiencies of selected E2s in an in vitro ubiquitination assay. UbcH5c and UbcH7 were both found to catalyse the ubiquitination of YB-1 in conjuction with RBBP6, whereas Ubc13 was not. Finally, we show using NMR that two single-point mutations of the RING finger-like domain are sufficient to abolish homodimerisation of the domain. These will be used in future studies to investigate the requirement for homodimerisation on the ubiquitination activity of RBBP6.en_US
dc.description.countrySouth Africa
dc.identifier.urihttps://hdl.handle.net/10566/13517
dc.language.isoenen_US
dc.publisherUniversity of the Western Capeen_US
dc.rights.holderUniversity of the Western Capeen_US
dc.subjectRBBP6en_US
dc.subjectYB-1en_US
dc.subjectInteractionen_US
dc.subjectRINGen_US
dc.subject15N-HSQCen_US
dc.subjectNMRen_US
dc.subjectYeasten_US
dc.subject2-hybriden_US
dc.subjectCo-immunoprecipitationen_US
dc.subjectHomodimerisationen_US
dc.subjectUbiquitinationen_US
dc.titleStructural characterisation of the interaction between RBBP6 and the multifunctional protein YB-1en_US
dc.typeThesisen_US

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Philosophiae Doctor - PhD (Biotechnology)