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Principal Investigator: Jacob O Brunkard
Organization: UNIVERSITY OF WISCONSIN-MADISON
Fiscal Year: 2024
Award: $320,956
Funding agency: National Institute of General Medical Sciences
PROJECT SUMMARY
TARGET OF RAPAMYCIN (TOR) is a deeply conserved protein kinase that regulates eukaryotic
metabolism. TOR senses and integrates upstream signals, especially nutrient availability, to coordinate
metabolism and promote growth only when conditions are favorable. TOR dysregulation causes or
contributes to a broad range of human diseases, including cancers, age-related health disorders, and
metabolic disorders, which are the major causes of morbidity and mortality in the United States.
Therefore, a major goal for biomedical research is to develop therapeutic treatments that specifically
target components of the TOR signaling network without broadly disrupting metabolism and
homeostasis in healthy cells that rely on TOR. Recently, there have been significant advances to that
goal with the discovery of several putative amino acid sensors for TOR. Conflicting reports about the
relative contributions, importance, and molecular mechanisms of these sensors have stymied these
advances, however. This project uses an innovative approach to bring fresh perspective to these
ongoing debates by shifting focus to the other major eukaryotic lineage, plants.
In my lab’s ongoing work to elucidate the TOR signaling network in plants, I discovered a novel amino
acid sensor for TOR, an aminoacyl tRNA synthetase (aaRS). This aaRS is necessary to maintain TOR
activity and sufficient to stimulate TOR in plant cells. Using a combination of biochemical, molecular,
genetic, and systems-level approaches, I propose to precisely define how the aaRS activates TOR in
plant cells through three independent aims. In Aim 1, I propose to mutate key enzymatic residues and
structural features of the aaRS to determine the molecular features it requires to activate TOR. In Aim
2, I propose to map the signal transduction pathway mediating aaRS-TOR activation using robust
orthogonal interactomic approaches. Putative signal transduction components will then be validated
using reciprocal assays and functional genetics to comprehensively define how aaRS-TOR interactors
contribute to TOR regulation. In Aim 3, I propose to establish the selective sensitivity of TOR for specific
amino acids and determine whether the aaRS is a bona fide amino acid sensor for TOR.
Taken together, these three aims will define the molecular mechanisms underlying the putative amino
acid-aaRS-TOR signaling axis and open new directions for future research on metabolic regulation in
eukaryotes. Moreover, this pathway will serve as a model for understanding how tRNA synthetases
have evolved functions beyond translation in signal transduction pathways and illuminate how the
complex TOR signaling network evolved to integrate diverse physiological cues in humans. Long-term,
our findings will make significant contributions to a major goal of contemporary biomedical research:
fine-tuning TOR signaling networks to improve and lengthen healthy human lifespans.
Terms: <ATP-protein phosphotransferase><Affinity Chromatography><Amino Acids><Amino Acyl T RNA Synthetases><Amino Acyl-tRNA Ligases><Amino Acyl-tRNA Synthetases><Aminoacyl Transfer RNA Synthetase><Aminoacyl-tRNA Synthetase><Arabidopsis><Assay><Autoregulation><Binding><Bioassay><Biochemical><Biologic Models><Biological Assay><Biological Models><Biology><Biomedical Research><Cancers><Catalysis><Cell Body><Cell Communication and Signaling><Cell Signaling><Cells><Complex><Cues><Development><Disease><Disorder><EIF-2 alpha><EIF-2alpha><EIF-2α><Eukaryota><Eukaryote><Evolution><Foundations><Future><Generalized Growth><Genetic><Genetic Alteration><Genetic Change><Genetic defect><Genomics><Goals><Growth><Growth and Development><Growth and Development function><Health><Heart><Homeostasis><Human><In Vitro><Intermediary Metabolism><Intracellular Communication and Signaling><Investigation><Kinase Family Gene><Kinases><Knowledge><Label><Leu-tRNA><Leucine><Malignant Neoplasms><Malignant Tumor><Maps><Mediating><Mediation><Metabolic><Metabolic Diseases><Metabolic Disorder><Metabolic Processes><Metabolism><Methodology><Methods><Model System><Modeling><Modern Man><Molecular><Molecular Interaction><Morbidity><Morbidity - disease rate><Mutate><Mutation><Negotiating><Negotiation><Nitrogen><Nutrient availability><Outcome><Pathway interactions><Phosphotransferase Gene><Phosphotransferases><Physiologic><Physiological><Physiological Homeostasis><Physiology><Phytonutrients><Plant Nutrients><Plant-based nutrients><Plants><Protein Kinase><Proteins><Proteomics><Rapamune><Rapamycin><Regulation><Reporting><Reproducibility><Role><Signal Pathway><Signal Transduction><Signal Transduction Pathway><Signal Transduction Systems><Signaling><Sirolimus><Solid><System><Techniques><Testing><Therapeutic><Thesaurismosis><Tissue Growth><Transfer RNA><Transfer RNA Synthetase><Translations><Transphosphorylases><Triplet Codon-Amino Acid Adaptor><Two Hybrid><United States><Variant><Variation><Work><Yeast One Hybrid System><Yeast One/Two-Hybrid System><affinity purification><age associated><age correlated><age dependent><age linked><age related><age specific><alpha Subunit Eukaryotic Initiation Factor 2><aminoacid><aminoacid tRNA ligase><biological signal transduction><detection of nutrient><developmental><experiment><experimental research><experimental study><experiments><genetic approach><genetic strategy><genome mutation><glycogen synthase a kinase><human disease><hydroxyalkyl protein kinase><improved><in vivo><innovate><innovation><innovative><leucine-tRNA><leucyl-tRNA><life span><lifespan><mRNA Translation><malignancy><metabolism disorder><metabolism measurement><metabolomics><metabonomics><mortality><neoplasm/cancer><novel><nutrient sensing><ontogeny><pathway><perception of nutrients><phosphorylase b kinase kinase><protein protein interaction><sensor><social role><tRNA><tRNA Synthetase><targeted drug therapy><targeted drug treatments><targeted therapeutic><targeted therapeutic agents><targeted therapy><targeted treatment><therapeutic target><transfer Ribonucleic acids><translation><yeast two hybrid system>