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EASTBIO: Investigating the assembly and trafficking of dynein motor complexes using an in vivo model


About This PhD Project

Project Description

In this project, the student will use state of the art imaging (both fluorescence and electron microscopy) to investigate the dynein assembly and transport processes. This will utilise Drosophila as a powerful model for a structural and functional analysis of motor complex assembly in vivo.

Amelia Shoemark is an expert in using electron micrscopy to characterise the molecular-scale defects in cilium structure in PCD patients, for both diagnostic and research purposes [1]. Andrew Jarman has pioneered the use of Drosophila as a model organism for analysing motile cilium defects and motor function, including the use of genetics, whole organism imaging, and proteomics [2,3]. In this project, the student will receive training that combines these different expertises, based in both Edinburgh and Dundee. The student will initially build a molecular model of Drosophila motile cilium structure using electron tomography (the first time this will be attempted). This will provide the baseline for tomography analysis of ciliary defects in mutants of a range of Drosophila genes orthologous to human PCD genes (recently characterised and assembled in the Jarman lab [3]). Characterisation of the ultrastructural defects will allow the student to propose mechanisms of protein complex assembly that can be explored by analysis of data on proteome changes in these mutants, as well as immunofluorescent microscopy analysis of motor assembly. In particular, the student will build on recent bioimaging work in the Jarman lab that uses SNAP-tagged motor protein subunits to follow real-time dynein assembly processes in live tissues by fluorescence microscopy. Overall, the student will gain a training in complementary bioimaging techniques, cellular and whole-organism analysis, as well as the manipulation and analysis of extensive proteomic datasets.

1. Burgoyne et al. Characterizing the ultrastructure of primary ciliary dyskinesia transposition defect using electron tomography. Cytoskeleton 2014 (https://doi.org/10.1002/cm.21171)
2. zur Lage et al. Ciliary dynein motor preassembly is regulated by Wdr92 in association with HSP90 co-chaperone, R2TP. J. Cell Biol. 2018 (https://doi.org/10.1083/jcb.201709026)
3. zur Lage et al. Survey of the ciliary motility machinery of Drosophila sperm and ciliated mechanosensory neurons reveals unexpected cell-type specific variations: a model for motile ciliopathies. Front. Genet. 2019 DOI:10.3389/fgene.2019.00024

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