Functional Characterization of Apical Kinases in the Cellular Stress Response

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My research program aims to understand how cells respond to different types of stress. I am particularly interested in dissecting the role of kinases that initiate stress response pathways and the underlying mechanisms. Our current focus is on the evolutionary conserved apical kinase ATM. Organisms must be able to respond to cellular stress resulting from internal and external origins to ensure their survival. Important responses include the regulation of countermeasures, cell cycle, metabolism, and for multicellular organisms apoptosis. Enzymatic posttranslational modifications, such as protein phosphorylation, enable rapid signal transduction and have evolved as efficient control mechanisms. Not surprisingly therefore, two apical protein kinases of stress response pathways are conserved among eukaryotes: The yeast kinases Tel1 and Mec1 share homology and functional similarity with the mammalian kinases ATM and ATR. Primarily investigated for the role these kinases have in the response to DNA damage, increasing evidence supports their role in the surveillance also of other types of insults such as oxidative, proteotoxic or mechanical stress. Having studied the role of ATM and ATR in the genotoxic stress response for many years, I propose to establish a program in the lab that uses novel approaches to expand our understanding of signal transduction pathways following cellular stress, including from different sources. We first will focus on ATM (Ataxia Telangiectasia Mutated) and in the future expand our studies to the related kinase ATR (Ataxia Telangiectasia and Rad3-related). Furthermore, based on the findings in the next five years, the program will continue studying the mechanisms of ATM regulation and the role of binding partners in determining the activation of kinases by cellular stress. To functionally characterize the role of ATM in the cellular stress response of various nature I propose: in Objective 1 to elucidate mechanisms of ATM activation by cellular stress by investigating stress-induced changes in the protein interactome by TurboID combined with cell fractionation; in Objective 2 to study the spatiotemporal dynamics of ATM localization after cellular stress by life cell imaging and static high resolution microscopy. The use of state-of-the arts techniques (including unpublished methods) and equipment will enable previously unattainable information on the regulation of ATM. Besides advancing the field, the research program will thereby also train HQP with a unique set of skills. IMPACT: Mechanistic insights into how cells respond to stress, whether arising from intrinsic or environmental factors threatening homeostasis, are fundamental to our understanding of life. Better knowledge of stress response pathways has also important implications for applied research, including the generation of induced pluripotent stem cells and tissue engineering, but also common lab techniques like tissue culture.