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Mechanisms for the selective clearance of chemically damaged proteins in mammalian cells

Project description

Uncovering the impact of nePTMs on protein turnover

Inside living cells, proteins are continuously exposed to reactive metabolites that introduce non-enzymatic post-translational modifications (nePTMs), such as oxidation, carbonylation, glycation, or carbamylation. nePTMs can irreversibly block critical interaction sites and pose a threat to proteome integrity. nePTM-bearing proteins are selectively degraded in healthy cells by an unknown mechanism that recognises and eliminates them. The EU-funded PRO-CLEAR project will study the impact of nePTMs on protein turnover and explore the underlying molecular mechanisms. It will test the effects of nePTMs and identify the adducts that induce protein clearance. PRO-CLEAR project aims to uncover a novel pathway for the quality control of chemically damaged proteins that will form the basis for understanding its role in physiology and human diseases.

Objective

Cellular homeostasis and survival critically depend on the ability to remove misfolded and damaged proteins. In cells, proteins are continuously exposed to reactive metabolites which introduce modifications such as oxidation, carbonylation, glycation or carbamylation. Such non-enzymatic post-translational modifications (nePTM) can irreversibly block critical interaction surfaces or reactive sites and thus pose a constant threat to proteome integrity. In line with this view, levels of nePTM are increased in diseased and aged tissues. In contrast, nePTM-bearing proteins are selectively degraded in healthy cells, suggesting the existence of a so far unknown pathway that recognizes and eliminates nePTM-bearing proteins.

In this project, I will study the impact of nePTMs on protein turnover and explore the underlying molecular mechanisms. More specifically, I plan to (I) establish a reporter assay to monitor the degradation of proteins displaying individual amino acid modifications in human cells. This will allow to test effects of common nePTMs and to identify the precise chemical adducts that induce protein clearance. Aim (II) is to uncover the cellular machinery that mediates nePTM-triggered protein degradation. To this end, I will perform a genome-wide CRISPR screen for genes required to eliminate a fluorescent model substrate. Aim (III) of the project is to understand the mechanisms underlying the recognition and clearance of nePTMs. I will therefore characterize one newly identified mediator of nePTM-clearance in detail and test its ability to bind, degrade or modify nePTM-bearing proteins and will study the impact of its knockout on the turnover of endogenous proteins.

Together these experiments promise to uncover a novel pathway for the quality control of chemically damaged proteins which will form the basis for understanding its role in physiology and human disease.

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Programme(s)

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Topic(s)

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Funding Scheme

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MSCA-IF - Marie Skłodowska-Curie Individual Fellowships (IF)

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Call for proposal

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(opens in new window) H2020-MSCA-IF-2019

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Coordinator

EIDGENOESSISCHE TECHNISCHE HOCHSCHULE ZUERICH
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.

€ 191 149,44
Address
Raemistrasse 101
8092 Zuerich
Switzerland

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Region
Schweiz/Suisse/Svizzera Zürich Zürich
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.

€ 191 149,44
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