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Title: Expression, molecular interactions and functions of Ruk, a novel adaptor protein
Author: Luke, C. T.
Awarding Body: University of Edinburgh
Current Institution: University of Edinburgh
Date of Award: 2005
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In this work I confirmed the Ruk gene structure using phage library analysis, which showed that it encompasses 320 kb and contains 24 exons with introns that are relatively large (³l00kb). There are 5 promoters found in Ruk and differential usage of these promoters together with alternative splicing generate 12 types of Ruk mRNA, which are for the majority developmentally down-regulated; the exceptions are Rukt and Rukh2 which are upregulated in adult testis. The 12 transcripts encode 7 different proteins and the only domain that is common for all of them is the coiled-coil. Therefore, the coiled-coil was targeted for conditional inactivation using the Cre loxP system. The tissue specific inactivation of Ruk dimerisation will help to elucidate the function of the individual isoforms depending on which tissues are targeted. GST pulldown and co-immunoprecipitation studies have shown that isoforms are able to interact with one another through various domains. Similar techniques have been used to elucidate the fine mechanism of the interaction between Ruk proteins and the p85α regulatory subunit of PI3-kinase. Ruk isoforms that were expressed in mammalian cells were shown to localize to vesicular structures, probably late endosomes, suggesting that certain domains interacting with membrane bound proteins cause the entire protein to be translocated to the endosomal membrane.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available