“Remodelling of Proteome Expression and Turnover during Cell Transformation”
Professor Angus Lamond FRS FRSE FMedSci is Professor of Biochemistry in the School of Life Sciences, Centre for Gene Regulation and Expression at the University of Dundee in Scotland. Before moving to Dundee in 1995 Angus was a group leader at the European Molecular Biology Laboratory in Heidelberg, where he started using mass spectrometry based proteomics techniques. Angus’ group study gene expression and the functional organization of mammalian cell nuclei, using a strategy that combines quantitative mass spectrometry (MS) based proteomics and live cell fluorescence imaging (see www.Lamondlab.com). The Lamond group have developed proteomic methods for studying the dynamics of protein localization, turnover and protein-protein interactions. They have created a software project – Peptracker – aimed at the efficient integration, analysis and open access sharing of multiple large proteomics datasets (see www.peptracker.com/epd). The current focus of the Lamond group is to identify and characterize gene regulatory mechanisms relevant to disease in both human cells and model organisms.
We have used a ‘Multidimensional Proteomics’ approach (see Larance and Lamond, Nat Rev. Mol. Cell Biol. 2015) to conduct an in-depth, quantitative analysis of the change in the proteomics landscape of healthy human epithelial cells that are induced to transform in culture by activation of the tyrosine kinase activity of the v-Src oncoprotein. We have characterized the proteome of untransformed MCF10A cells to a depth of ~14,000 proteins, detecting over 350,000 separate peptides with a mean coverage of >25 peptides per protein. These cells were induced to transform in culture by activation of v-Src, resulting in major phenotypic changes within 48-72 hours. The transformed cells show many of the hallmark phenotypes of metastatic cancer cells seen in the clinic, including changes in morphology, loss of contact inhibition and increases in both migration and invasiveness. We have quantitated by label free MS analysis the response kinetics for the entire MCF10A proteome, measuring seven time points, in biological triplicate, spanning 1-72 hours post v-Src activation. We have also used pulsed SILAC to measure, in biological triplicate, the rates of protein synthesis, degradation and turnover in both healthy and v-Src transformed MCF10A human epithelial cells. All of these data have been integrated in a searchable, online database featuring user-friendly, interactive data exploration tools (see www.peptracker.com/epd). I will present an overview of the resulting data, which provide a detailed insight at a systems level into the molecular events and remodeling of metabolism and gene expression that accompany the formation of a cancer cell. I will also relate these data from a human cellular model to clinical data and healthcare records.