Hasperhoven, Aliek (2022) Mapping the molecular makeup of the inner kinetochore in the slime mould Dictyostelium discoideum. Master's Research Project 1, Biomolecular Sciences.
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Abstract
Eukaryotic cell division relies on the kinetochore, a multi-protein structure at the centromere of each chromosome, to facilitate the separation of duplicated chromosomes into daughter cells. This process involves the attachment of microtubules to the kinetochore, which ensures even distribution of sister chromatids between daughter cells. Dysregulation of this process can lead to cancer. In most eukaryotes, including humans, the inner kinetochore comprises CENP-A, the Constitutive Centromere Associated Network (CCAN), and the Chromosomal Passenger Complex (CPC). However, the kinetochore of Dictyostelium discoideum exhibits unique characteristics, such as the absence of CENP-C in the CCAN and the presence of a Survivin-like gene with one helix and no chromatin-binding domains in the CPC. To better understand the kinetochore in this organism, we used gene-tagging, mass spectrometry analysis, and high-resolution imaging. We constructed a BioID variant miniTurbo vector to enable proximity-based biotinylation with nuclear proteins, which was used for biotin pulldowns. The study confirmed the presence and interactions of several inner kinetochore proteins but did not provide a clear explanation for the absence of CENP-C. The function and localization of the Survivin ortholog remain unknown, and further optimization of the methods is necessary to fully address these questions.
| Item Type: | Thesis (Master's Research Project 1) |
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| Supervisor name: | Tromer, E.C. and Kortholt, A. |
| Degree programme: | Biomolecular Sciences |
| Thesis type: | Master's Research Project 1 |
| Language: | English |
| Date Deposited: | 17 Apr 2023 10:03 |
| Last Modified: | 17 Apr 2023 10:03 |
| URI: | https://fse.studenttheses.ub.rug.nl/id/eprint/29599 |
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