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Molecular interconnections between transcription, translation and DNA repair in bacteria

Project description

The interplay of gene expression processes

The complex interplay between bacterial gene expression processes remains poorly understood. Key mechanisms like transcription, translation, and DNA repair often operate in parallel, yet little is known about how they are functionally coupled. In this context, the ERC-funded RNP5D project seeks to address this by investigating how these processes interact at the molecular level. Using advanced techniques in single-molecule biophysics, biochemistry, and structural biology, the project will reconstitute transcription-translation systems and DNA repair pathways in vitro and in vivo. By integrating these systems, the project aims to uncover how these critical cellular processes cooperate or compete, providing new insights into gene expression regulation and setting a new standard for studying complex biological systems.

Objective

This proposal brings together the fields of bacterial transcription, translation and DNA repair with state-of-the art single-molecule biophysics in vitro and in vivo, biochemistry and structural biology to address a central but understudied question in biology: how are different macromolecular processes interconnected and functionally coupled? I aim to elucidate this by investigating the crosstalk between central processes in bacterial gene expression: transcription, translation and DNA repair.
First, to understand what determines whether transcription is coupled to translation, we will reconstitute the complete transcription-translation system in vitro and image the process in real-time using cutting-edge multi-color single-molecule fluorescence experiments. We will support our dynamic single-molecule data with structural information from cryo-electron tomography. In order to verify that our in vitro data also have physiological relevance, we will electroporate reconstituted and labelled RNAP/ribosome complexes into live E. coli cells for multi-color single-molecule imaging also in vivo.
Second, we will reconstitute the transcription-coupled DNA nucleoside excision repair pathway in vitro. Integrating multi-color single-molecule fluorescence with optical tweezer assays, will allow us to resolve a controversial point on how transcription-coupled DNA repair starts, comparing the established concept of Mfd-initiation with the newly proposed UvrD/RNAP-mediated lesion recognition.
Finally, we will develop single-molecule experiments for integrating all three processes, specifically transcription, translation and DNA repair, into a single system to quantify cooperativity and competition between individual components and processes.
Overall, this proposal will provide quantitative and mechanistic understanding of how central processes to gene expression are functionally interconnected, setting a new benchmark for examining complex biological systems in context.

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HORIZON-ERC - HORIZON ERC Grants

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Call for proposal

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(opens in new window) ERC-2024-COG

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Host institution

EUROPEAN MOLECULAR BIOLOGY LABORATORY
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.

€ 2 614 460,00
Address
Meyerhofstrasse 1
69117 Heidelberg
Germany

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Region
Baden-Württemberg Karlsruhe Heidelberg, Stadtkreis
Activity type
Research Organisations
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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.

€ 2 614 460,00

Beneficiaries (1)

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