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Tension of ENDOmembranes maintained by TORC1

Project description

TOR proteins in the regulation of vacuolar membrane tension

The serine/threonine-protein kinase, TOR, is the target of the anti-fungal natural product rapamycin and exists in two complexes: TORC1 and TORC2. TORC1 is directly inhibited by rapamycin and plays important roles in biomass accumulation while TORC2 functions to maintain tension homeostasis of the plasma membrane. Researchers of the EU-funded TENDO project will develop novel chemical probes and use a broad methodological arsenal to investigate if and how TORC1 analogously senses and regulates tension of endomembranes. Moreover, they will study if and how TORC2 is regulated through formation of a helix-like structure upon glucose depletion as is the case for TORC1. This work will reveal new mechanisms of TOR regulation that may translate into therapeutic interventions.

Objective

The target of the bacterial macrolide rapamycin, TOR, is a ser/thr protein kinase that assembles into two distinct protein complexes, conserved from yeast to human, we named TORC1 and TORC2. TORC1 is directly bound and inhibited by rapamycin and studies with rapamycin have revealed that TORC1 plays a central role in coupling nutrient cues to biomass synthesis and turnover. The lack of a specific inhibitor for TORC2 has made the study of this complex much more challenging. We overcame this challenge by solving the structure of yeast TORC2 which revealed why it is insensitive to rapamycin and enabled us to create a rapamycin-sensitive TORC2 variant. We also developed two small molecules, one that dissipates plasma membrane (PM) tension and the other that serves as a biosensor of PM tension. With this suite of chemical-biology tools we confirmed that TORC2 functions in a mechanotransduction pathway to maintain tension homeostasis of the PM. Concurrently, solving the structure of TORC1 revealed that its activity is regulated via assembly into a huge, inactive helix which we named a TOROID – TORC1 Organized in an Inactive Domain. In this grant, was ask if these major advances are transferable; i.e. can lessons learned regarding TORC2 be applied to TORC1, and vice versa? Our major aim is to determine if and how TORC1 regulates vacuolar membrane (VM) tension. To this end, we will develop novel chemical probes to monitor VM tension and we will use genetic screens, quantitative phosphoproteomics, in vitro assays, high-throughput compound screens, STORM and FRAP imaging, and state-of-the-art cryo-EM to learn how TORC1 senses and regulates VM tension. Our other aim, prompted by our TOROID discovery, is to solve the TOROID-like structure that TORC2 forms upon glucose depletion. This work will reveal new mechanisms in growth control, and details in TORC1 and TORC2 regulation that may inform future therapeutic interventions for these medically relevant signalling complexes.

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Topic(s)

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ERC-ADG - Advanced Grant

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Call for proposal

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(opens in new window) ERC-2018-ADG

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Host institution

UNIVERSITE DE GENEVE
Net EU contribution

Net EU financial contribution. The sum of money that the participant receives, deducted by the EU contribution to its linked third party. It considers the distribution of the EU financial contribution between direct beneficiaries of the project and other types of participants, like third-party participants.

€ 2 257 546,00
Address
RUE DU GENERAL DUFOUR 24
1211 Geneve
Switzerland

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Region
Schweiz/Suisse/Svizzera Région lémanique Genève
Activity type
Higher or Secondary Education Establishments
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Total cost

The total costs incurred by this organisation to participate in the project, including direct and indirect costs. This amount is a subset of the overall project budget.

€ 2 257 546,00

Beneficiaries (1)

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