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Comprehending Prokaryotic Argonaute Systems

Project description

Role of Argonaute proteins in prokaryotes

Argonaute proteins are components of the RNA-induced gene silencing known as RNA interference. They bind different classes of small non-coding RNAs, which guide them to specific targets, leading to mRNA cleavage or translation inhibition. Yet, their evolutionary origin lies in prokaryotes. The EU-funded COMPASS project will map the function of unexplored prokaryotic Argonaute protein (pAgo) systems, in which pAgos associate with auxiliary proteins. The hypothesis is that pAgos bind exogenous DNA sequences, resulting in the recruitment of the auxiliary proteins and fulfilling completely novel roles ranging from stimulating horizontal gene transfer to triggering programmed cell death. The project results will enable the repurposing of programmable pAgo systems for the development of genetic tools.

Objective

Eukaryotic Argonaute proteins are known for their central role in RNA interference pathways. Yet, the evolutionary origin of Argonaute proteins lies in prokaryotes, where other proteins essential for RNA interference are absent. Therefore, prokaryotic Argonaute proteins (pAgos) must have distinct ancestral functions. Although a handful of closely related pAgos interfere with exogenous DNA invaders such as plasmids, pAgos are extremely diverse in terms of sequence conservation and domain architecture. In addition, many pAgos genetically associate with various putative enzyme domains, which suggests that they are functionally interdependent. As such, different pAgos must rely on distinct mechanism and are expected to fulfil a range of different roles. Therefore, the function of most pAgos remains completely unknown.

The COMPASS project will map the function of unexplored pAgo systems, in which pAgos associate with auxiliary proteins. I hypothesize that in these systems, pAgo binds exogenous DNA sequences in a guide-dependent manner. This can result in the recruitment and/or activation of the auxiliary proteins. As pAgo-associated auxiliary proteins are homologous to proteins involved in DNA recombination, NAD+ turnover, or protein deacetylation, these pAgo systems are expected to fulfil completely novel roles ranging from stimulating horizontal gene transfer to triggering programmed cell death.

The uncharacterized roles of these pAgo systems and the mechanisms underlying their functionality will be elucidated by a multidisciplinary approach combining microbiology, protein biochemistry, and X-ray crystallography techniques. Not only will the results facilitate a deeper understanding of the evolutionary diversification of pAgos, it will also enable the repurposing of programmable pAgo systems for the development of genetic tools that facilitate guide sequence-directed DNA recombination and high-sensitivity detection of target DNA sequences.

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ERC-STG - Starting Grant

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(opens in new window) ERC-2020-STG

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Host institution

WAGENINGEN UNIVERSITY
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.

€ 1 496 463,00
Address
DROEVENDAALSESTEEG 4
6708 PB Wageningen
Netherlands

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Region
Oost-Nederland Gelderland Veluwe
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.

€ 1 496 463,00

Beneficiaries (1)

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