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In vivo characterization of Hsp104 variants in Saccharomyces cerevisiae

Cell stress, caused by misfolded or aggregated proteins or exposure to certain environmental stressors, activates stress detectors and selected analysis processes within the cell, resulting intranscriptional regulation and synthesis of factors that influence appropriate folding or removal of misaligned proteins to recover proteostasis. Yeasts are given a sophisticated and interconnected system to restore from the disruption of proteostasis, which involves molecular chaperones and protein degradation pathways as significant participants. Hsp104 is a stress response protein that, through an unknown mechanism, stimulates the reactivation of heatdamaged proteins in yeast. The aim of this thesis study is to learn more about Hsp104 function and location in young and old yeast cells. On the one hand, we aim to see how Hsp104 wildtype vs. mutant forms are distributed in young vs. elderly yeast cells. As a negative control, we employed the mutant variant of this protein. We also wanted to stress Hsp104 wildtype and mutant versions to see whether there are any variations in behaviour or protein levels. All research was carried out in yeast, with biochemical tests and high-throughput technologies like flow cytometry. Hsp104GFP signal were increased in the nucleus of Hsp104 wildtype cells with the increasing of cellular age. As well as, after heatshock treatment the Hsp104GFP aggregation was raised in Hsp104 wildtype and mutant forms. The results data demonstrated that Hsp104 protein levels were increased with cellular age and heatshock treatment enhanced the Hsp104 aggregation in Hsp104 wildtype and mutant forms.

Identiferoai:union.ndltd.org:UPSALLA1/oai:DiVA.org:his-20187
Date January 2021
CreatorsRahman, Md. Mominur
PublisherHögskolan i Skövde, Institutionen för biovetenskap
Source SetsDiVA Archive at Upsalla University
LanguageEnglish
Detected LanguageEnglish
TypeStudent thesis, info:eu-repo/semantics/bachelorThesis, text
Formatapplication/pdf
Rightsinfo:eu-repo/semantics/openAccess

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