Structural studies of eukaryotic protein synthesis factor complexes by Cryo EM

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Molecular recognition drives almost all biological reactions. This project aims to uncover structural details of the interactions between protein factors critical for translation in all eukaryotic cells. The specific translation initiation factors are the G protein eIF2 and two regulators called eIF2B and eIF5. eIF2 is required to bind initiator tRNA to ribosomes. Its GTP/GDP status is controlled by eIF2B and eIF5. Phosphorylation of eIF2 at a specific serine residue inhibits eIF2B activity by forming a non-productive eIF2:eIF2B complex. This mechanism of translational control is conserved across all studied eukaryotes and is important within diverse settings including developmental, metabolism and long-term memory formation. eIF2 and eIF2B are both large multi-subunit complexes, and mutations within these factors cause neurological diseases. We aim to deconstruct the mechanism of action of these complexes, and their regulation processes. The structure and interactions between purified translation factor complexes will be elucidated by state-of-the-art molecular cryo electron microscopy techniques.This project incorporates cutting-edge biophysical analysis methods with innovative data analysis. It will blend modern precision molecular biology techniques with biochemistry and yeast genetics. The project involves protein expression and purification, molecular biology, electron microscopy, 3D structure analysis and molecular model building.This project will require 3D visualisation of data and statistical information to reliably interpret small differences in different protein complexes. We aim to get information on protein complex motion and dynamic information, as well as individual structures. Novel visualisation methods will be developed, as well as new quantitative 3Dimage analysis methods.