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Mechanism of Type 9 Secretion: an unusual protein transport system involved in pathogenesis and motility

Project description

Pathogenic protein secretion in bacteria

Bacteria have evolved complex mechanisms for exporting virulence factors across their envelope to target cells. Until recently, six different protein-based secretion systems (I-VI) were known in Gram-negative bacteria. The EU-funded SecNine project aims to characterise the newly discovered type IX secretion system of the Gram-negative phylum Bacteroidota. Researchers are working to identify the implicated proteins, determine their molecular structure and delineate their interaction during the transport process. The generated information will provide fundamental knowledge on a novel pathogenic mechanism of bacteria that are responsible for severe periodontal disease.

Objective

The recently-discovered Type IX Secretion System (T9SS) is a protein transport pathway that exports proteins across the outer membrane of Gram-negative of the phylum Bacteroidetes. It is an essential pathogenicity determinant in severe periodontal disease and in the major bacterial diseases of farmed fish. It is also required for the unique Bacteroidete gliding motility. The components required to catalyse T9SS export are still not fully defined, but include at least 18 authenticated proteins. Possible functions can currently only be assigned to some of these T9SS components. However, it is clear that the pathway involves a number of interacting protein complexes.
Our current understanding of the T9SS is superficial and lacking in molecular level structural and mechanistic detail. In a landmark recent study we have identified and structurally characterized the multi-protein outer membrane protein conducting channel of the T9SS. Our ambitious goal is to apply similar state-of-the-art experimental methods to systematically characterise the full T9SS pathway.
We will systematically characterise the component protein complexes that make up the T9SS by:
- Defining their composition through co-purification from native sources.
- Determining their molecular structures by cryo-EM.
- Assessing their dynamic interactions through live cell fluorescence imaging, biochemical experiments, and structural characterization.
In addition, we will use single molecule tracking methods to follow substrate molecules through the steps of the T9SS transport cycle in real time in live bacterial cells and apply this methodology to elucidate T9SS mechanism.

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Programme(s)

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Topic(s)

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Funding Scheme

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ERC-ADG - Advanced Grant

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Call for proposal

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(opens in new window) ERC-2018-ADG

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Host institution

THE CHANCELLOR, MASTERS AND SCHOLARS OF THE UNIVERSITY OF OXFORD
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.

€ 2 245 365,00
Address
WELLINGTON SQUARE UNIVERSITY OFFICES
OX1 2JD Oxford
United Kingdom

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Region
South East (England) Berkshire, Buckinghamshire and Oxfordshire Oxfordshire
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.

€ 2 245 365,00

Beneficiaries (1)

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