Use this URL to cite or link to this record in EThOS: https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.589263
Title: A whole genome RNAi screen to identify novel promoters of PINK1/Parkin-mediated mitophagy
Author: Ivatt, R. M.
Awarding Body: University of Sheffield
Current Institution: University of Sheffield
Date of Award: 2013
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Abstract:
The PINK1/Parkin pathway is genetically linked to recessive forms of Parkinson's disease (PD) and plays a central role in the maintenance of mitochondrial homeostasis. Following toxification with the membrane uncoupler CCCP, PINK1, a serine/threonine kinase, becomes stabilised on the outer membrane of damaged mitochondria. This stabilisation is necessary for the translocation of Parkin, an E3- ubiquitin ligase, to mitochondria prior to their removal via mitophagy. In order to identify novel pathway components upstream of Parkin translocation, we performed a genomewide RNAi screen using Drosophila S2R+ cells. Upon completion, we validated primary screen hits with multiple rounds of secondary screening, assessing a range of mitochondrial homeostatic processes. In follow-up screens, corresponding human orthologs were assessed for their ability to influence Parkin translocation and mitophagy in HeLa cells, demonstrating conserved pathway function. Human screen hits selected for in depth analysis included several genes acting in a common lipogenesis pathway. These are the transcription factor and master pathway regulator SREBF1, the Skp1-Cul1-F-box E3-ligase protein FBXW7 and the serine/threonine kinase GSK3A. Follow-up analyses of these genes suggest a role for SREBPdependent lipid synthesis in PINK1/Parkin-mediated mitophagy, likely acting upstream of PINK1. Importantly, SREBF1 has recently been identified as a risk locus for sporadic forms of PD (Do et al, 2011). Hence, this study reveals a novel mechanistic link between familial- and sporadic causes of PD, with mitochondrial dysfunction at its centre.
Supervisor: Whitworth, Alex Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID: uk.bl.ethos.589263  DOI: Not available
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