The recent recognition that numerous proteins and protein regions perform functions while lacking folded globular structure has opened a new perspective on protein structure-function relationships. This project focuses on understanding the role of unstructured proteins and protein domains in protein-protein interactions. The main objective of this project is to determine whether intrinsic protein disorder is involved or possibly even required for protein-protein interactions involving multiple binding partners. The hypothesis that will be tested is that proteins with multiple binding partners interact with their partners via natively disordered regions/domains. A combined approach using computational and experimental methods is devised to achieve these goals. First, a computational analysis that includes disorder predictions on proteins with multiple interacting partners from the C. elegans interactome will be performed. In addition, the algorithm for predicting disorder-to-order transition mutations in these proteins will be developed. Second, prediction-guided and domain-guided approaches will be used to select the targets for experimental verification. Third, site-directed and deletion mutagenesis of predicted disordered regions followed by a high-throughput yeast two-hybrid (Y2H) screening using known binding partners (derived from the large-scale C. elegans interactome mapping project) will be carried out. Finally, a database that contains all C. elegans interaction domains together with their respective partners defined in this study will be assembled and disseminated.
The interdisciplinary nature of this project will advance knowledge, stimulate scientific exchange, and integrate research and education across two scientific fields - biology and computer science. Data analysis and the development of a database of interacting protein domains will provide excellent research and training opportunities to high school students, graduate students, and postdoctoral research associates in the new and largely unexplored area of intrinsic protein disorder.
