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Towards the discovery of cellular RNA-binding proteins with master regulatory roles in virus infection

Project description

Cellular master regulator proteins in RNA viruses infection

RNA viruses have RNA as their genetic material which plays a central role during infection. The viruses use cellular RNA-binding proteins (RBPs) to support infection. Host cells also employ RBPs to combat viruses. The EU-funded vRNP-capture project aims to discover the repertoire of cellular RBPs with master regulatory roles in RNA virus infection. A viral RNA interactome capture, a novel method developed in the host laboratory, will allow the elucidation of viral ribonucleoprotein composition with unprecedented depth and specificity. The technology employs pulse-labeling of viral RNA with a photoactivatable nucleotide analogue, UV crosslinking, viral RNA purification and quantitative proteomics. It will enable the discovery of cellular RBPs with master regulatory roles in infection as potential targets for broad-spectrum antiviral therapies.

Objective

RNA is a central molecule for RNA virRNA is a central molecule for RNA virus infection. However, viral genomes encode very few proteins that are able to interact with viral RNA. Hence, viruses co-opt cellular RNA-binding proteins (RBPs) to support infection. The host cell also employs RBPs to combat viruses through the recognition of unusual signatures present viral RNAs. Despite these critical roles, the complement of cellular RBPs involved in infection remains largely unknown. This research programme aims to discover comprehensively and unbiasedly the repertoire of cellular RBPs endowed with master regulatory roles in virus infection. To achieve this, we will exploit a novel method developed in my laboratory, named viral RNA interactome capture (vRIC), that allows the elucidation of viral ribonucleoprotein (RNP) composition with unprecedented depth and specificity. It employs pulse-labelling of viral RNA with a photoactivatable nucleotide analogue, UV crosslinking, viral RNA purification with antisense probes and quantitative proteomics. (Aim 1) I hypothesise that cellular RBPs that interact with a broad-range of viral RNAs are likely endowed with master regulatory roles in infection. To test this, I will apply vRIC to cells infected with different RNA viruses to discover RBPs that are shared across viral RNPs. Moreover, we will employ interferon α to investigate if the engagement of these RBPs with viral RNA is regulated by the antiviral state. (Aim 2) To test if these broad-spectrum RBPs display master regulatory roles in infection, I will apply a novel functional screen using genetically modified cells and a broad library of fluorescent viruses. (Aim 3) We will then decipher the molecular mechanisms underpinning the ability of these RBPs to support or restrict infection. In summary, this innovative research programme will discover cellular RBPs with master regulatory roles in infection with great potential as targets for broad-spectrum antiviral therapies.

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Programme(s)

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Topic(s)

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Funding Scheme

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ERC-COG - Consolidator Grant

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Call for proposal

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(opens in new window) ERC-2020-COG

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Host institution

UNIVERSITY OF GLASGOW
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.

€ 1 999 916,00
Address
UNIVERSITY AVENUE
G12 8QQ Glasgow
United Kingdom

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Region
Scotland West Central Scotland Glasgow City
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.

€ 1 999 916,00

Beneficiaries (1)

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