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Tuning the Tracks: Resolving the interplay between microtubule lattice conformation and kinesin-1 selectivity using cryo-EM

Objective

Kinesin-1 transports cellular cargos along microtubules as part of neuronal transport. The localisation of kinesin-1 in neurons is tightly controlled to ensure the coordinated delivery of specific cargoes to the regions where they are required. One mechanism of control is through microtubule regulation, but the details of this are not well understood. It has recently been shown that microtubule lattice spacing influences kinesin-1 localisation in U2OS cells. However, the role lattice spacing plays in neuronal transport is unclear. This project aims to investigate features of kinesin-1 based neuronal transport on multiple scales. First, this project aims to use cryo-electron microscopy (cryo-EM) to elucidate how kinesin-1 binds to specific microtubule subsets. I will resolve the high-resolution cryo-EM structure of a dimeric kinesin-1 construct (stableMARK) bound to microtubules with different lattice states. This will explain its lattice specificity and help uncover the mechanics of kinesin-1 walking. Secondly, I will map microtubule lattice spacing in neurons using in situ cryo-electron tomography to establish the lattice diversity within this complex microtubule network. Thirdly, I will use cryo-correlative light electron microscopy and stableMARK to determine if lattice spacing influences kinesin-1 localisation in neurons. This project will answer key biological questions on cytoskeletal transport, which will be applicable to other microtubule-based motors and microtubule associated proteins. Defects in kinesin-1 based transport have been implicated in multiple neuropathies, such as Alzheimer’s disease, therefore further understanding of how neuronal transport is controlled will help uncover the mechanism of these diseases.

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Programme(s)

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Topic(s)

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HORIZON-TMA-MSCA-PF-EF - HORIZON TMA MSCA Postdoctoral Fellowships - European Fellowships

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Call for proposal

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(opens in new window) HORIZON-MSCA-2023-PF-01

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Coordinator

UNIVERSITEIT UTRECHT
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.

€ 187 624,32
Address
HEIDELBERGLAAN 8
3584 CS Utrecht
Netherlands

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Activity type
Higher or Secondary Education Establishments
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Total cost

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