[SCItalk] Using the bioUb strategy to elucidate the molecular mechanisms of Angelman Syndrome
University of the Basque Country (UPV/EHU), Spain
Angelman syndrome is a cognitive disorder caused by the lack of function in the brain of UBE3A. Also known as E6-AP, this ubiquitin E3 ligase had previously been studied in proliferative cell culture, but nothing was known about its ubiquitination substrates in neuronal cells. Application of the bioUb strategy to both Drosophila and mice model systems allowed us to identify several of its neuronal substrates. The UBE3A-mediated regulation of the proteasome (several of its subunits) has a large relevance for Angelman Syndrome, since it could explain the complex symptoms of this monogenic disorder. Further identification of the ubiquitination sites and chain types on UBE3A substrates opens a gateway for functional studies on the relevance and role of these ubiquitination events. Practical examples will be presented on various approaches to answer classical key questions on the ubiquitin field: Which proteins does a given E3 ubiquitinate? How do I confirm that my protein of interest is indeed deubiquitinated by a candidate DUB enzyme? Is my protein mono- or polyubiquitinated? How do I quantitatively measure changes on chain linkages of my protein upon differential conditions? How do I discover the E3 ligase that ubiquitinates my protein? Finally, I will also present work in progress for a database of the human DUB ubiquitome.
Dr. Mayor´s PhD research (University of Cambridge, UK, 2003) focused on protein dynamics and structure and resulted on the first detailed analysis ever done of an ultrafast folding protein. The main experiments from his thesis were published in Nature in 2003; being cited 520 times since. He then moved to the Gurdon Institute in Cambridge, UK, where he was awarded with a Royal Society Dorothy Hodgkin Fellowship. At this time Dr. Mayor developed a novel method to isolate and identify, in a tissue specific manner, the neuronal targets of the ubiquitination machinery within a living organism. Dr. Mayor moved then to Spain (CIC bioGUNE, Bilbao) in 2009, where he started his own lab. In 2011 he was awarded a March of Dimes Basil O ́Connor Fellowship to start research on the molecular mechanisms affected in Angelman Syndrome. At this time, the bioUb strategy was licensed to be used on a stable cell line, and he also signed a contract with local company Histocell and UK-based Ubiquigent for commercialization of MEF cells derived from the bioUb mice. At the end of 2014, Dr. Mayor moved to the Biochemistry Department of the Basque Country University (UPV/EHU), gaining access to a much better setup of mass spectrometry equipment. His lab is currently focused on understanding the molecular mechanisms that govern Angelman Syndrome.
Host: Jose Luis Rosa - Cell Signaling and Bone Biology group