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Structural and biochemical studies on the regulation of transcription during elongation

Project description

RNA polymerase and transcription: biochemical and structural investigations

Gene expression begins with DNA transcription, during which the DNA message is copied onto a new molecule, messenger RNA, by the universally conserved enzyme RNA polymerase (RNAP). This step is essential to the healthy functioning of organisms and is implicated in a growing number of diseases. Despite its importance and considerable progress, the process is still incompletely understood. The ERC-funded TRANSREG project will use advanced biochemical and structural approaches combined with studies of the dynamics of RNAP to address critical questions concerning RNAP dynamics during processes that regulate transcription. Although focused on bacterial RNAP, insight and experience could pave the way to similar studies in more complex organisms.

Objective

During the first step of gene expression, a universally conserved enzyme called RNA Polymerase (RNAP) transcribes RNA from a DNA template in every cell. Transcription is a fundamental, yet incompletely understood process. Because of its central role, the bacterial transcription machinery is also a prime drug target. Crystal structures of RNAP core as well as RNAP elongation complexes (ECs) have revolutionized our understanding of transcription. This groundwork paved the way to address more complex questions regarding the regulation of transcription. Transcriptional regulation impacts every aspect of biology including a growing list of human diseases. Transcriptional pausing, a temporary interruption of transcription during elongation, plays a major role in regulating gene expression in pro- and eukaryotes. A full and mechanistic understanding is crucial and requires a combination of structural and biochemical studies. This is where the proposed work will contribute.

Our goal is to understand how transcription factors modulate RNAP during the elongation phase.

Biochemical and structural approaches (X-ray crystallography and single particle cryo-EM) will be used to gain detailed insights on the regulation of transcription by protein factors and non-coding RNAs (ncRNAs). To complement the results, an experimental system will be established using Fluorescence Resonance Energy Transfer (FRET) to study the dynamics of RNAP. This system will allow us to address questions concerning RNAP dynamics during processes that regulate transcription.

Specifically, we propose 3 aims:
1.: Investigate the role of transcription elongation factors that modulate RNAP pause behavior and couple transcription and translation
2.: Study the role of RNAs that regulate transcription
3.: Establish an experimental system to study RNAP dynamics using FRET

The proposed work will lay the groundwork and prepare the team to address similar questions in higher organisms in the future.

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Programme(s)

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Topic(s)

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Funding Scheme

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ERC-STG - Starting Grant

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Call for proposal

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(opens in new window) ERC-2015-STG

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Host institution

CENTRE EUROPEEN DE RECHERCHE EN BIOLOGIE ET MEDECINE
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.

€ 1 500 000,00
Address
Rue Laurent Fries 1
67404 Illkirch Graffenstaden
France

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Region
Grand Est Alsace Bas-Rhin
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.

€ 1 500 000,00

Beneficiaries (1)

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