Alhammad, Yousef (2024) Characterisation of peroxisomes in the fission Yeast Schizosaccharomyces pombe and slime mold Dictyostelium discoideum. PhD thesis, University of Sheffield.
Abstract
Peroxisome is a compartment that is found in most eukaryotic organisms' cells. It has several crucial roles, such as fatty acid beta (β) oxidation and hydrogen peroxide (H2O2) detoxification. It contains many essential enzymes, including oxidase and catalase, and has several metabolic and non-metabolic pathways, depending on the environment and the organisms within its cells. This study investigates the role of peroxisomes in two organisms, S. pombe and D. discoideum.
Although S. pombe is a well-studied yeast, there is only one study of this yeast that has focused on peroxisomes. This study offers a few crucial observations, including that S. pombe contains peroxisomes, that GFP containing a well-characterized PTS1 (SKL) is efficiently imported, and that peroxisome numbers increase in cells grown on a fatty acid as the sole carbon source, suggesting a role for peroxisomes in fatty acid degradation.
The starting point in my research was initially a bioinformatics screen. This screening recognized the enzymes imported into peroxisomes based on the presence of a potential peroxisomal targeting signal. A few proteins were found. However, the low number of proteins with a classical PTS might be the result of different targeting signals that are not recognized by our bioinformatics parameters. Indeed, in other organisms, there are proteins without PTS1 that still use Pex5 for import. The first example is S. cerevisiae Acyl-CoA oxidase. In a global yeast two-hybrid screen, S. pombe Pex5 was found to bind S. pombe Str3 and Lys3. Consequently, we think that there is conserved targeting of a peroxisomal protein lacking a PTS1 and PTS2 imported into the peroxisome by Pex5. One of these is the Str3 case. Interestingly, proteins involved in peroxisomal fatty acid β -oxidation are absent from the S. pombe genome, casting doubt on the conclusions from the previous study and explaining the low number of potential peroxisomal enzymes.
In D. discoideum, this study investigates the dynamic regulation of peroxisome numbers in response to growth conditions and identifies peroxisomal import and contents through a proximity labeling approach (BioID). Overall, this study sheds light on the roles and regulation of peroxisomes in these two organisms.
Metadata
Supervisors: | Jason, King and Ewald, Hettema and Donald, Watts |
---|---|
Keywords: | Peroxisome; Schizosaccharomyces pombe; Dictyostelium discoideum; fatty acid beta (β) oxidation; proximity labeling approach (BioID). |
Awarding institution: | University of Sheffield |
Academic Units: | The University of Sheffield > Faculty of Science (Sheffield) The University of Sheffield > Faculty of Science (Sheffield) > Molecular Biology and Biotechnology (Sheffield) |
Depositing User: | Mr Yousef Mohammed A. Alhammad |
Date Deposited: | 13 Feb 2024 10:39 |
Last Modified: | 13 Feb 2024 10:39 |
Open Archives Initiative ID (OAI ID): | oai:etheses.whiterose.ac.uk:34269 |
Download
Final eThesis - complete (pdf)
Filename: Yousef_Alhammad_thesis_Final_version .docx
Licence:
This work is licensed under a Creative Commons Attribution NonCommercial NoDerivatives 4.0 International License
Export
Statistics
You do not need to contact us to get a copy of this thesis. Please use the 'Download' link(s) above to get a copy.
You can contact us about this thesis. If you need to make a general enquiry, please see the Contact us page.