Journal Article

Systems-level effects of allosteric perturbations to a model molecular switch

Kortemme, Tanja, Ollikainen, Noah, Zhang, Yang, Perica, Tina, Braberg, Hannes, Krogan, Nevan J., Mathy, Christopher J. P., Xu, Jiewei, Jang, Gwendolyn Μ., Lambright, David G., Kaake, Robyn, Swaney, Danielle L., Kelly, Mark J. S.

2021

Overview

Molecular switch proteins whose cycling between states is controlled by opposing regulators 1 , 2 are central to biological signal transduction. As switch proteins function within highly connected interaction networks 3 , the fundamental question arises of how functional specificity is achieved when different processes share common regulators. Here we show that functional specificity of the small GTPase switch protein Gsp1 in Saccharomyces cerevisiae (the homologue of the human protein RAN) 4 is linked to differential sensitivity of biological processes to different kinetics of the Gsp1 (RAN) switch cycle. We make 55 targeted point mutations to individual protein interaction interfaces of Gsp1 (RAN) and show through quantitative genetic 5 and physical interaction mapping that Gsp1 (RAN) interface perturbations have widespread cellular consequences. Contrary to expectation, the cellular effects of the interface mutations group by their biophysical effects on kinetic parameters of the GTPase switch cycle and not by the targeted interfaces. Instead, we show that interface mutations allosterically tune the GTPase cycle kinetics. These results suggest a model in which protein partner binding, or post-translational modifications at distal sites, could act as allosteric regulators of GTPase switching. Similar mechanisms may underlie regulation by other GTPases, and other biological switches. Furthermore, our integrative platform to determine the quantitative consequences of molecular perturbations may help to explain the effects of disease mutations that target central molecular switches. Interface mutations in the GTPase switch protein Gsp1 (the yeast homologue of human RAN) allosterically affect the kinetics of the switch cycle, revealing a systems-level mechanism of multi-specificity.

• Publisher: Nature Publishing Group UK,Nature Publishing Group
• Subject: 38/47 / 38/70 / 631/1647/2067 / 631/45/612/1232 / 631/553/2711 / 631/57/2272/2273 / 631/92/360 / 82/58 / 82/6 / 82/80 / 82/83 / Allosteric properties / Allosteric Regulation - genetics / Binding sites / Binding Sites - genetics / Biological activity / Catalytic Domain - genetics / Cell cycle / Genes / GTPase-Activating Proteins - metabolism / Guanine Nucleotide Exchange Factors - metabolism / Guanosine triphosphatases / Guanosine Triphosphate - metabolism / Homology / Humanities and Social Sciences / Interfaces / Kinases / Kinetics / Molecular machines / Monomeric GTP-Binding Proteins - genetics / Monomeric GTP-Binding Proteins - metabolism / multidisciplinary / Mutation / Nuclear Proteins - genetics / Nuclear Proteins - metabolism / Perturbation / Point Mutation / Post-translation / Protein Binding - genetics / Proteins / Quantitative genetics / Regulation / Saccharomyces cerevisiae - enzymology / Saccharomyces cerevisiae - genetics / Saccharomyces cerevisiae Proteins - genetics / Saccharomyces cerevisiae Proteins - metabolism / Science / Science (multidisciplinary) / Signal transduction / Switches