Principal Supervisor: Prof Corinne SmithLink opens in a new window, Life Sciences

Co-supervisor: Dr Nikola Chmel, Chemistry

PhD project title: Dissecting the mechanism of clathrin-coated vesicle formation

University of Registration: University of Warwick

Project outline:

Clathrin-mediated endocytosis is a fascinating mechanical phenomenon that drives the selective internalisation of molecules into cells. Nutrient uptake, synaptic vesicle recycling, signalling, determination of cell polarity and development all rely on endocytic mechanisms. In disease, viral and bacterial pathogens exploit endocytosis to gain entry into cells and malfunctions lead to tumour formation, neurodegeneration and heart disease. In order to work properly, clathrin-mediated endocytosis requires accurate and timely assembly of a clathrin lattice and coordination with a network of more than 20 adaptor proteins to form a coated vesicle which will be able to select molecules from the outside of the cell for delivery to specific destinations. Our aim is to understand how the proteins involved in the network of clathrin and its adaptor proteins interact to achieve clathrin-coated vesicle formation.

In this project you will use biophysical and structural biology techniques to visualise adaptor proteins binding to clathrin cages and to analyse the dynamics of those interactions. These techniques include high resolution cryo-electron microscopy, small angle neutron scattering, dynamic light scattering, time-resolved fluorescence anisotropy and surface plasmon resonance (SPR, Biacore). By combining results from this wide range of approaches we will be able to investigate how clathrin-adaptor interactions result in formation of a functional coated vesicle network. Access will be provided to the Gatan K2 Summit direct detector and Jeol 2200FS 200kV transmission electron microscope at the Advanced Bioimaging Research Technology Platform, the Titan Krios microscope available in Leicester through the Midlands Regional Cryo-EM Facility and excellent facilities for biophysical analysis available within Warwick School of Life Sciences. This is a fabulous opportunity to apply cutting edge techniques to discovering how clathrin and its adaptor proteins drive clathrin mediated endocytosis.

 

References:

Smith SM, Smith CJ. Capturing the mechanics of clathrin-mediated endocytosis. Curr Opin Struct Biol. (2022) 75:102427

Smith SM, Larocque G, Wood KM, Morris KL, Roseman AM, Sessions RB, Royle SJ, Smith CJ. Multi-modal adaptor-clathrin contacts drive coated vesicle assembly. EMBO J. (2021) 40(19):e108795.

Morris KL, Jones JR, Halebian H, Wu S, Baker M, Armache J, Ibarra AA, Sessions RB, Cameron AD, Cheng Y, Smith CJ. Cryo-EM of multiple cage architectures reveals a universal mode of clathrin self assembly. Nature Structural and Molecular Biology (2019) 26(10):890-898.

 

 

BBSRC Strategic Research Priority: Understanding the rules of life – Structural Biology, and Integrated Understanding of Health - Ageing, and Diet and Health.

 

Techniques that will be undertaken during the project:

Kinetic analysis using light scattering, fluorescence and single molecule methods

Small-angle neutron scattering

High resolution electron microscopy

Protein mutagenesis, expression and purification

Image analysis of large data sets

3D imaging of structural data

 

Contact: Prof Corinne SmithLink opens in a new window