Grant: Project Grant

Turning personal human genome sequences into actionable information for treatment or genetic engineering requires understanding how individual and groups of mutations modify the activity proteins quantitatively. One of the most critical impacts of mutations on proteins is changing how they organize each other in space and time through their physical associations. Such changes underlie many human diseases. Thanks to sustained CIHR support, we have developed approaches based on genetic perturbations (amino acid variants, copy-number variation) to quantify interactions among proteins in living cells and how mutations impact their strength and specificity. We now request funding to assess how entire interactomes respond to mutations. Crucial interactions for eukaryotic cell biology are between SH3 domains and their binding motifs. These interactions, for instance, play essential roles in ancient, fundamental processes such as endocytosis. Here, we will harness our tools to dissect all of the SH3-directed protein-protein interactions of a biomedical cell model at the level of single residues within proteins. Our work will provide a roadmap for interpreting mutations affecting the binding of proteins and the specificity of these interactions at the network levels. Our results will represent the most comprehensive data on the impact of mutations on the entire cellular network in living cells. This work will help guide the interpretation of genetic variation in human cell homologous protein interaction networks.