About the Project
You will be part of our collaborative working environment and have access to outstanding shared facilities such as microscopy and proteomics. Throughout your year, you will develop an advanced level of knowledge on your topic of interest as well as the ability to perform independent research in the topic area. Alongside basic science training in experimental design, data handling and research ethics, we will help you to develop skills in critical assessment and communication. This will be supported by workshops in scientific writing, presentation skills, ethics, laboratory safety, statistics, public engagement and optional applied bioinformatics.
The period of study is one year full-time or two years part-time research, which includes two months to write up the thesis. Please apply via the UCAS postgraduate application form: https://digital.ucas.com/courses/details?coursePrimaryId=c735d826-42b6-ca1f-50db-2a3ac6f68718
Protein S-acylation is a poorly understood fatty acid based post-translational modification of proteins, yet affects half of all membrane proteins and plays a role in every membrane associated process in plants. We have shown the S-acylation is key to fundamental aspects of plant biology including cellulose synthase function, pathogen perception, hormone signalling and water homeostasis during environmental stress. Knowledge of S-acylation is therefore crucial to understanding the basic biology of plant function and is key to many aspects of plant biology required for mitigating climate change, water shortages or boosting crop yield.
We recently found that S-acylation state can change in response to stimuli, is enzymatic and has profound effects on protein function. This places S-acylation alongside phosphorylation and ubiquitination in terms of regulatory importance but the mechanism underlying how changes in protein S-acylation state occurs is still unclear. We have identified enzymes that add S-acyl groups to proteins and have a shortlist of candidate de-S-acylating enzymes able to remove S-acyl groups; the basis for a regulatory cycle therefore exists. Using model S-acylated proteins involved in pathogen perception, hormone responses and water stress mitigation this project aims to 1) validate these enzyme candidates and 2) develop strategies to monitor dynamic S-acylation of proteins in plants.
1. Validation of candidate de-S-acylating enzymes
a. Express cloned candidate de-S-acylating enzymes in plants and validate their activity and sensitivity to inhibitors of de-S-acylation using chemical biology tools.
b. Validated candidates will be tested for their ability to remove S-acyl groups from model proteins in plants.
2. Monitoring dynamic S-acylation changes
a. In mammalian systems it is possible to monitor S-acylation turnover by feeding cells fatty acid analogues containing a tag that allows for visualisation or enrichment (a.k.a. metabolic labelling). Plants, being autotrophic, do not take up fatty acids from the environment with high efficiency. It is therefore necessary to develop alternative routes to achieve metabolic labelling using these fatty acid analogues. You will test different strategies including the use of protoplasts, expression of animal/fungal fatty acid transporters in plants and the use of carrier compounds to promote uptake of fatty acids into plant cells. The successful strategy will then be used to monitor S-acylation turnover at the whole proteome level, and at the level of the individual proteins described in part 1.
This project will provide training in plant and protein biochemistry/molecular biology, chemical biology, methods development and quantitative analysis. Potential applicants are encouraged to contact Piers Hemsley ([Email Address Removed]) for further information and informal discussion about the project.
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