Supervisor Project : Establishing the role of SUMO protease SENP6 in regulating SUMO chain length
Ron Hay is a Wellcome Trust Senior Investigator and a fellow of the Royal Society, the Royal Society of Edinburgh, the Academy of Medical Sciences, Academia Europaea and is a member of the European Molecular Biology Organisation. In 2012 Ron was awarded the Novartis Medal and Prize of the Biochemical Society. He is currently Chair of Molecular Biology in the University of Dundee and is part of the Centre for Gene Regulation and Expression. Ron is also an honorary member of The MRC Protein Phosphorylation and Ubiquitylation Unit.
Loss of function studies with essential genes such as SENP6 are limited to conditional knock out of the gene or mRNA depletion with siRNA. The drawback with both of these approaches is that substantial depletion of the gene product takes at least 48 hours and with a very stable protein even longer. To circumvent these problems we have recently employed AID technology in live worms to deplete SUMO E3 ligase GEI17 in less than 60 minutes. Our first objective is therefore to use CRISPR/cas9 technology to engineer cell lines expressing the auxin responsive plant protein TIR1 and endogenous SENP6 tagged with GFP-AID. Using these cell lines we will determine the critical phases of the cell cycle where SENP6 depletion has deleterious consequences for passage through mitosis. Similar cell lines also expressing 6His T90K SUMO2 will allow us to realise the objective of isolating proteins containing extended SUMO chains and mapping sites of SUMO modification and thus define physiological substrates of SENP6.The final objective is to determine the basis for selective cleavage of SUMO2 chains by SENP6. In collaboration with the lab of Huib Ovaa, this will be accomplished by chemically linking SENP6 to diSUMO and determining the structure of the complex by X-ray crystallography.