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Compartment-specific proteostasis

Kalsbeek, Robin (2021) Compartment-specific proteostasis. Research Project 1, Biomedical Sciences.


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Protein quality control (PQC) is of vital importance of the cell. Since the various cellular compartments are confronted with distinct challenges, it is natural that the different compartments have evolved distinct PQC pathways. While much work has focused on the PQC pathways of the cytosol and ER, the PQC networks inside the nucleus are to this date the more elusive. The thermolabile protein firefly luciferase offers many benefits to study protein quality control and is therefore a very popular tool. However, the studies that have used firefly luciferase in the past were often lacking in comparing their findings to standardized control proteins. As such, in the present study we have put together a standardized, modularized library of plasmids that express proteins with varying thermodynamic properties, that localize to different compartments, and are fluorescently tagged. We show that nuclear localization signal (NLS)-Firefly luciferase-GFP localizes to the nucleolus upon heat stress, but that NLS-Renilla luciferase-GFP forms peri-nucleolar foci upon heat stress. We show that the use of a nuclear export signal (NES) improves cytoplasmic localization, but also causes accumulation at the aggresome. We have generated plasmids that express glutathione S-transferase (GST) fusion proteins that are low expressed and stable, and double fluorophore proteins that are highly expressed and stable. Lastly, we have made plasmids that express targeted versions of poly- glutamine (poly-Q) d

Item Type: Thesis (Research Project 1)
Supervisor name: Klaips, C.L.
Degree programme: Biomedical Sciences
Thesis type: Research Project 1
Language: English
Date Deposited: 27 Jul 2021 14:03
Last Modified: 27 Jul 2021 14:03

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