MRC DiMeN Doctoral Training Partnership: Investigating the role of Golga3 cleavage during SARS-CoV-2 infection

Description

SARS-CoV-2 is the causative agent of the COVID-19 pandemic, responsible for >600 million infections and over six and a half million deaths worldwide by September 2022. The virus produces the proteins it needs for replication by the proteolytic cleavage of two polyproteins by the viral Mpro and PLP proteases. But, these proteases also attack host proteins.

The Emmott lab and collaborators used quantitative mass spectrometry to identify 14 cellular proteins cleaved by these viral proteases in SARS-CoV-2-infected cells (Meyer et al. 2021). We demonstrated that these target proteins were essential for efficient SARS- CoV-2 replication. One hit was GOLGA3 (Golgin subfamily A, member 3), which was cleaved by SARS-CoV-2 Mpro.

GOLGA3 is ubiquitously expressed and maintains Golgi apparatus structure, subcellular trafficking and apoptosis. We hypothesise that GOLGA3 cleavage plays a role in the remodeling of cytoplasmic membranes during SARS-CoV-2 infection, as well as the secretion of assembled SARS-CoV-2 virions.

This project seeks to characterize the localization, interactions and cleavage kinetics of GOLGA3 to better understand the role of this protein in normal cell biology, as well as to provide a mechanistic understanding of how GOLGA3 cleavage contributes to SARS-CoV-2 replication. This work will synergise with newly MRC-funded research in the Emmott lab on cellular targets of viral proteases.

This studentship covers both ‘wet-lab’ and computational training primarily in the areas on molecular/cellular biology, virology and proteomics. In this project, you will use LC-MS/MS-based proteomic assays and microscopy to study viral proteolytic cleavage and protein interactions of GOLGA3. The student will also employ molecular biology approaches including lentivirus-mediated CRISPR-Cas9 mutagenesis to generate endogenous GOLGA3 mutant cell lines to test the impact of different GOLGA3 modifications on the replication of SARS-CoV-2 and other human coronaviruses (229E, OC43, NL63). You will spend time working with the Ungar lab at the University of York, to test the impact of these mutations on Golgi function and export, and with the Pat Eyers lab at Liverpool, testing the impact of inhibitors on Golga3 function. You will also gain training in data analysis in R/Matlab.

You will primarily be working with Dr Ed Emmott (Twitter: @edemmott, emmottlab.org) at the Centre for Proteome Research, University of Liverpool. You will also be working with Prof. Dani Ungar (York, Ungar, Prof Daniel - Biology, University of York) to study the impact of Golga3 cleavage on Golgi function, and with Prof. Pat Eyers (Liverpool , Twitter: @Pseudoenzyme , Patrick Eyers - Institute of Systems, Molecular and Integrative Biology - University of Liverpool) to study the impact of inhibitors on the extensively-phosphorylated Golga3 protein. You are very strongly encouraged to email the lead supervisor Ed Emmott () to talk through the project before you make your application.

Benefits of being in the DiMeN DTP:

This project is part of the Discovery Medicine North Doctoral Training Partnership (DiMeN DTP), a diverse community of PhD students across the North of England researching the major health problems facing the world today. Our partner institutions (Universities of Leeds, Liverpool, Newcastle, York and Sheffield) are internationally recognised as centres of research excellence and can offer you access to state-of the-art facilities to deliver high impact research.

We are very proud of our student-centred ethos and committed to supporting you throughout your PhD. As part of the DTP, we offer bespoke training in key skills sought after in early career researchers, as well as opportunities to broaden your career horizons in a range of non-academic sectors.

Being funded by the MRC means you can access additional funding for research placements, international training opportunities or internships in science policy, science communication and beyond. See how our current DiMeN students have benefited from this funding here: https://www.dimen.org.uk/blog

Further information on the programme and how to apply can be found on our website: https://www.dimen.org.uk/how-to-apply

Availability

Open to students worldwide

Funding information

Funded studentship

Studentships are fully funded by the Medical Research Council (MRC) for 4yrs. Funding will cover tuition fees, stipend and project costs. We also aim to support the most outstanding applicants from outside the UK and are able to offer a limited number of full studentships to international applicants. Please read additional guidance here: View Website
Studentships commence: 1st October 2023
Good luck!
 

 

Supervisors

References

Meyer et al. (2021) Characterising proteolysis during SARS-CoV-2 infection identifies viral cleavage sites and cellular targets with therapeutic potential. Nature Comms. https://www.nature.com/articles/s41467-021-25796-w
Emmott et al. (2019) Polyprotein processing and intermolecular interactions within the viral replication complex spatially and temporally control norovirus protease activity. J. Biol. Chem.
https://www.sciencedirect.com/science/article/pii/S0021925820418319?via%3Dihub
Emmott et al. (2017) Norovirus-mediated modification of the translational landscape via virus and host-induced cleavage of initiation factors. Mol. Cell. Proteomics.
https://www.sciencedirect.com/science/article/pii/S0021925820475532?via%3Dihub