High-content functional genomic screening to identify novel regulators of the PINK1-Parkin pathway

• Published in: Methods in enzymology Vol. 547; p. 1
• Main Authors: Ng, Andy Cheuk-Him, Baird, Stephen D, Screaton, Robert A
• Format: Journal Article
• Language: English
• Published: United States 2014
• Subjects: Algorithms; ATPase Inhibitory Protein; Electronic Data Processing; Genomics - instrumentation; Genomics - methods; HeLa Cells; High-Throughput Nucleotide Sequencing - instrumentation; High-Throughput Nucleotide Sequencing - methods; Humans; Protein Kinases - genetics; Protein Kinases - metabolism; Proteins - genetics; Proteins - metabolism; RNA Interference; RNA, Small Interfering; Ubiquitin-Protein Ligases - genetics; Ubiquitin-Protein Ligases - metabolism; F1-Fo ATP synthase; RNA interference screen; Mitochondria; PINK1; IF1; ATPIF1; Mitophagy; PARK2
• ISSN: 1557-7988
• DOI: 10.1016/B978-0-12-801415-8.00001-1

PINK1/PARK6 and Parkin/PARK2 are amongst the most commonly mutated genes associated with recessive forms of familial Parkinson's disease. Recent evidence indicates that the proteins they encode, PINK1 and Parkin, function in the same pathway to mediate the selective autophagic clearance of dysfunctional mitochondria. Upon mitochondrial damage, PINK1 is stabilized on the outer mitochondrial membrane where it phosphorylates ubiquitin, generating a signal for the recruitment and activation of Parkin. However, key mechanistic questions still exist regarding Parkin recruitment, including whether or not other factors are required for the PINK1 and Parkin pathway. We describe a method below using high-throughput RNA interference technology to interrogate the genome for novel components of the PINK1 and Parkin pathway.