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 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 to study the impact of Golga3 cleavage on Golgi function, supported by Prof. Pat Eyers (Liverpool). Please email the primary supervisor (firstname.lastname@example.org) for more details
Open to students worldwide
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.
Emmott et al. (2017) Norovirus-mediated modification of the translational landscape via virus and host-induced cleavage of initiation factors. Mol. Cell. Proteomics.