Study of the yeast cytosolic Hsp70-system in protein homeostasis and life span regulation - Hsp70-mutant yeast cells with the misfolding marker protein guk1-7-GFP, the nucleolar marker Sik1-RFP and nuclear staining with DAPI imaged before, directly after, and during recovery from heat stress

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.

Fluorescence microscopy of live yeast cells at mid-exponential growth before stress, directly after 30 minutes of heat shock and 40 minutes after heat shock. The yeast is grown in complete synthetic medium lacking uracil with 2 % glucose as carbon source. All strains produce the aggregate marker protein guk1-7-GFP and carry a plasmid expressing SIK1-RFP under the control of the GPD-promoter.
The dataset was collected through fluorescence microscopy.
The image files are provided in Carl Zeiss Image format (.czi).

本研究旨在解答如下科学问题：热休克蛋白70（Hsp70）类分子伴侣（molecular chaperones）的哪些功能，对于酵母维持标准复制寿命至关重要？
为解答该问题，我们在缺失Ssa1/2的酵母菌株中，利用Hsp70家族成员Ssa1、Ssa2及其旁系同源蛋白Ssa4的差异化功能，并通过异位手段提升了该菌株中Ssa4的蛋白生成量。本研究采集了多种实验条件下不同蛋白质聚集标记物的行为数据，以及其他类别分子伴侣的蛋白质相关数据。其中绝大多数数据来自宽场显微镜（widefield microscopy）拍摄的多通道显微图像（multichannel microscopy images），另有少量蛋白质提取物的蛋白质印迹（western blots）实验结果。
实验分别在应激前、热激30分钟后及热激40分钟后，对处于指数生长中期的活酵母细胞进行荧光显微镜成像。酵母培养于缺乏尿嘧啶的完全合成培养基中，以2%葡萄糖作为碳源。所有菌株均表达聚集标记蛋白guk1-7-GFP，并携带以GPD启动子调控表达SIK1-RFP的质粒。
本数据集通过荧光显微镜技术采集得到，图像文件采用蔡司图像格式（Carl Zeiss Image format，.czi）存储。