Study of the yeast cytosolic Hsp70-system in protein homeostasis and life span regulation - Timelapse microscopy of the misfolding protein guk1-7-GFP in recovery after heat stress in Hsp70- and Hsp104-mutant yeast strains

Our study aims to answer the question "Which functions of the Hsp70 class of molecular chaperones are essential for yeast to maintain a standard replicative life span?". To answer this question, we utilised the disparate functions of the Hsp70's Ssa1 and 2 and their paralog Ssa4 in a yeast strain that lacks Ssa1/2 but has an ectopically increased production of Ssa4. We have gathered data on the behaviour of several different markers for protein aggregation under different circumstances, as well as data on proteins from other classes of molecular chaperones. The bulk of the data is in the form of multichannel microscopy images from widefield microscopy, with a few sets of western blots of protein extracts. Timelapse microscopy of yeast cells during recovery after 30 minutes of heat shock. The yeast was cultivated in complete synthetic media with 2 % glucose as carbon source. All strains produce the misfolding protein guk1-7-GFP. The dataset was collected through timelapse microscopy of the same yeast cells for 2 hours. The image files are provided in Carl Zeiss Image format (.czi).本研究旨在解答以下问题：热休克蛋白70（Hsp70）家族分子伴侣（molecular chaperones）的哪些功能对酵母维持标准复制寿命至关重要？为解答该问题，本研究利用了Hsp70家族Ssa1、Ssa2及其旁系同源基因Ssa4的差异化功能，实验所用酵母菌株缺失Ssa1/2，但可异位高表达Ssa4。我们收集了多种蛋白质聚集标记物在不同条件下的行为数据，以及其他类别分子伴侣的相关蛋白数据。本数据集的主体数据为宽场显微镜（widefield microscopy）获取的多通道显微图像，辅以少量蛋白质提取物的免疫印迹实验结果。 本实验对经历30分钟热休克后恢复过程中的酵母细胞进行了延时显微成像。实验所用酵母在以2%葡萄糖为碳源的完全合成培养基中培养，所有菌株均表达错误折叠蛋白guk1-7-GFP。 本数据集通过对同一批酵母细胞进行2小时的延时显微成像获取。 图像文件以蔡司图像格式（Carl Zeiss Image Format，.czi）提供。