Study of the yeast cytosolic Hsp70-system in protein homeostasis and life span regulation

Creator/Principal investigator(s)

Rebecca Andersson - University of Gothenburg, Institute of Biomedicine, Department of Microbiology and Immunology orcid

Anna-Maria Eisele - University of Gothenburg, Institute of Biomedicine, Department of Microbiology and Immunology orcid

Sarah Hanzén - Cochlear Nordic AB orcid

Katarina Vielfort - Umeå University, Institute of Molecular Biology orcid

David Öling - AstraZeneca, Mölnlycke, Sweden, Discovery Biology, Discovery Sciences, Innovative Medicines and Early Development Biotech Unit orcid

Frederik Eisele - University of Gothenburg, Institute of Biomedicine orcid

Gustav Johansson - University of Gothenburg, Institute of Biomedicine orcid

Tobias Gustafsson - University of Massachusetts Medical School, Department of Biochemistry and Molecular Pharmacology orcid

Kristian Kvint - University of Gothenburg, Institute of Biomedicine, Department of Infectious Diseases orcid

Thomas Nyström - University of Gothenburg, Institute of Biomedicine, Department of Microbiology and Immunology orcid

Description

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.

Principal organisation

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Responsible department/unit

University of Gothenburg, Institute of Biomedicine, Department of Microbiology and Immunology

Creator/Principal investigator(s)

Rebecca Andersson - University of Gothenburg, Institute of Biomedicine, Department of Microbiology and Immunology orcid

Anna-Maria Eisele - University of Gothenburg, Institute of Biomedicine, Department of Microbiology and Immunology orcid

Sarah Hanzén - Cochlear Nordic AB orcid

Katarina Vielfort - Umeå University, Institute of Molecular Biology orcid

David Öling - AstraZeneca, Mölnlycke, Sweden, Discovery Biology, Discovery Sciences, Innovative Medicines and Early Development Biotech Unit orcid

Frederik Eisele - University of Gothenburg, Institute of Biomedicine orcid

Gustav Johansson - University of Gothenburg, Institute of Biomedicine orcid

Tobias Gustafsson - University of Massachusetts Medical School, Department of Biochemistry and Molecular Pharmacology orcid

Kristian Kvint - University of Gothenburg, Institute of Biomedicine, Department of Infectious Diseases orcid

Thomas Nyström - University of Gothenburg, Institute of Biomedicine, Department of Microbiology and Immunology orcid

Identifiers

SND-ID: 2020-36

Description

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.

Language

English

Time period(s) investigated

2012 — 2020

Unit of analysis

Population

Saccharomyces cerevisiae (Baker's yeast)

Study design

Experimental study

Preclinical study

Sampling procedure

Total universe/Complete enumeration

Funding

Knut and Alice Wallenberg Foundation

Publications

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Andersson R, Eisele-Bürger AM, Hanzén S, Vielfort K, Öling D, Eisele F, Johansson G, Gustafsson T, Kvint K, Nyström T. Differential role of cytosolic Hsp70s in longevity assurance and protein quality control. bioRxiv. 2020 Jun 29. Available from: https://www.biorxiv.org/content/10.1101/2020.06.25.170670v2.full
DOI: https://doi.org/10.1101/2020.06.25.170670

Andersson R, Eisele-Bürger AM, Hanzén S, Vielfort K, Öling D, Eisele F, Johansson G, Gustafsson T, Kvint K, Nyström T. Differential role of cytosolic Hsp70s in longevity assurance and protein quality control. PLoS Genetics. Accepted manuscript, 2020.
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Dataset 1

The misfolding protein marker gus1-3-GFP and the metacaspase Mca1-GFP during mid-exponential growth in yeast Hsp70-mutant yeast strains

Suggested citation

Rebecca Andersson, Sarah Hanzén, Katarina Vielfort, Thomas Nyström. University of Gothenburg, Institute of Biomedicine, Department of Microbiology and Immunology (2020). <em>The misfolding protein marker gus1-3-GFP and the metacaspase Mca1-GFP during mid-exponential growth in yeast Hsp70-mutant yeast strains</em>. Swedish National Data Service. Version 1. <a href="https://doi.org/10.5878/8wxm-8r38">https://doi.org/10.5878/8wxm-8r38</a>

Creator/Principal investigator(s)

Sarah Hanzén - Cochlear Nordic AB orcid

Katarina Vielfort - Umeå University, Institute of Molecular Biology orcid

Rebecca Andersson - University of Gothenburg, Institute of Biomedicine, Department of Microbiology and Immunology orcid

Thomas Nyström - University of Gothenburg, Institute of Biomedicine, Department of Microbiology and Immunology orcid

Description

Fluorescence microscopy of live yeast cells at mid-exponential growth. All strains produce the aggregate marker protein gus1-3-GFP or the metacaspase Mca1 tagged with GFP (Mca1-GFP)

The dataset was collected through fluorescence microscopy.
The image files are provided in Carl Zeiss Image format (.czi).

Data format / data structure

Still image

Data collection

Mode of collection: Biological tests

Time period(s) for data collection: 2017-05-29 — 2017-07-04

Data collector: University of Gothenburg

Source of the data: Research data, Biological samples

Dataset 2

The misfolding protein marker guk1-7-GFP before, during and after in recovery from heat shock stress in Hsp70-mutant yeast strains

Suggested citation

Rebecca Andersson, Thomas Nyström. University of Gothenburg, Institute of Biomedicine, Department of Microbiology and Immunology (2020). <em>The misfolding protein marker guk1-7-GFP before, during and after in recovery from heat shock stress in Hsp70-mutant yeast strains</em>. Swedish National Data Service. Version 1. <a href="https://doi.org/10.5878/8r0v-jk54">https://doi.org/10.5878/8r0v-jk54</a>

Creator/Principal investigator(s)

Rebecca Andersson - University of Gothenburg, Institute of Biomedicine, Department of Microbiology and Immunology orcid

Thomas Nyström - University of Gothenburg, Institute of Biomedicine, Department of Microbiology and Immunology orcid

Description

Fluorescence microscopy of live yeast cells at mid-exponential growth before stress, directly after 30 minutes of heat shock, and 20, 40 and 60 minutes after heat shock. The yeast is grown in complete synthetic medium with 2 % glucose as carbon source. All strains produce the aggregate marker protein guk1-7-GFP.
The dataset was collected through fluorescence microscopy.
The image files are provided in Carl Zeiss Image format (.czi).

Data format / data structure

Still image

Data collection

Mode of collection: Biological tests

Time period(s) for data collection: 2017-05-15 — 2017-05-26

Data collector: University of Gothenburg

Source of the data: Forskningsdata: Opublicerade, Biological samples

Dataset 3

Intracellular colocalisation of the chaperone Ssa4-GFP and the metacaspase Mca1-RFP before and after heat stress in a Hsp70-mutant yeast cell strain

Suggested citation

Rebecca Andersson, Katarina Vielfort, Thomas Nyström. University of Gothenburg, Institute of Biomedicine, Department of Microbiology and Immunology (2020). <em>Intracellular colocalisation of the chaperone Ssa4-GFP and the metacaspase Mca1-RFP before and after heat stress in a Hsp70-mutant yeast cell strain</em>. Swedish National Data Service. Version 1. <a href="https://doi.org/10.5878/hvf7-3j18">https://doi.org/10.5878/hvf7-3j18</a>

Creator/Principal investigator(s)

Katarina Vielfort - Umeå University, Institute of Molecular Biology orcid

Rebecca Andersson - University of Gothenburg, Institute of Biomedicine, Department of Microbiology and Immunology orcid

Thomas Nyström - University of Gothenburg, Institute of Biomedicine, Department of Microbiology and Immunology orcid

Description

Fluorescence microscopy of live yeast cells at mid-exponential growth before and after 30 minutes of heat shock. All strains produce the Hsp70 chaperone protein Ssa4 tagged with GFP (Ssa4-GFP) and the metacaspase Mca1 tagged with RFP (Mca1-RFP).
The dataset was collected through fluorescence microscopy.
The image files are provided in Carl Zeiss Image format (.czi).

Data format / data structure

Still image

Data collection

Mode of collection: Biological tests

Time period(s) for data collection: 2019-03-13 — 2019-03-14

Data collector: University of Gothenburg

Source of the data: Forskningsdata: Opublicerade, Biological samples

Dataset 4

The molecular chaperone GFP-Hsp104 before and after heat stress in Hsp70-mutant yeast strains

Suggested citation

Rebecca Andersson, Sarah Hanzén, Thomas Nyström. University of Gothenburg, Institute of Biomedicine, Department of Microbiology and Immunology (2020). <em>The molecular chaperone GFP-Hsp104 before and after heat stress in Hsp70-mutant yeast strains</em>. Swedish National Data Service. Version 1. <a href="https://doi.org/10.5878/gv2g-xc30">https://doi.org/10.5878/gv2g-xc30</a>

Creator/Principal investigator(s)

Sarah Hanzén - Cochlear Nordic AB orcid

Rebecca Andersson - University of Gothenburg, Institute of Biomedicine, Department of Microbiology and Immunology orcid

Thomas Nyström - University of Gothenburg, Institute of Biomedicine, Department of Microbiology and Immunology orcid

Description

Fluorescence microscopy of live yeast cells at mid-exponential growth and after 30 minutes of heat shock. The cells were grown in complete synthetic media lacking histidine (CSM-HIS) with 2 % galactose as carbon source. All strains carry a plasmid expressing GFP-HSP104 under the control of the GAL-promoter.

The dataset was collected through fluorescence microscopy.
The image files are provided in Carl Zeiss Image format (.czi).

Data format / data structure

Still image

Data collection

Mode of collection: Biological tests

Time period(s) for data collection: 2018-03-28 — 2018-03-29

Data collector: University of Gothenburg

Source of the data: Research data, Forskningsdata: Opublicerade, Biological samples

Dataset 5

Timelapse microscopy of the misfolding protein guk1-7-GFP in recovery after heat stress in Hsp70- and Hsp104-mutant yeast strains

Suggested citation

Rebecca Andersson, Sarah Hanzén, Thomas Nyström. University of Gothenburg, Institute of Biomedicine, Department of Microbiology and Immunology (2020). <em>Timelapse microscopy of the misfolding protein guk1-7-GFP in recovery after heat stress in Hsp70- and Hsp104-mutant yeast strains</em>. Swedish National Data Service. Version 1. <a href="https://doi.org/10.5878/wj4m-y925">https://doi.org/10.5878/wj4m-y925</a>

Creator/Principal investigator(s)

Sarah Hanzén - Cochlear Nordic AB orcid

Rebecca Andersson - University of Gothenburg, Institute of Biomedicine, Department of Microbiology and Immunology orcid

Thomas Nyström - University of Gothenburg, Institute of Biomedicine, Department of Microbiology and Immunology orcid

Description

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).

Data format / data structure

Still image

Data collection

Mode of collection: Biological tests

Time period(s) for data collection: 2019-03-19 — 2019-04-18

Data collector: University of Gothenburg

Source of the data: Forskningsdata: Opublicerade, Biological samples

Dataset 6

The molecular chaperone GFP-Hsp104 before and after heat stress in a Hsp70-mutant yeast strain with exogenous complementation of wildtype and chimaeric mutant alleles of yeast Hsp70-alleles

Suggested citation

Rebecca Andersson, Sarah Hanzén, Katarina Vielfort, Thomas Nyström. University of Gothenburg, Institute of Biomedicine, Department of Microbiology and Immunology (2020). <em>The molecular chaperone GFP-Hsp104 before and after heat stress in a Hsp70-mutant yeast strain with exogenous complementation of wildtype and chimaeric mutant alleles of yeast Hsp70-alleles</em>. Swedish National Data Service. Version 1. <a href="https://doi.org/10.5878/mez1-3z30">https://doi.org/10.5878/mez1-3z30</a>

Creator/Principal investigator(s)

Rebecca Andersson - University of Gothenburg, Institute of Biomedicine, Department of Microbiology and Immunology orcid

Sarah Hanzén - Cochlear Nordic AB orcid

Katarina Vielfort - Umeå University, Institute of Molecular Biology orcid

Thomas Nyström - University of Gothenburg, Institute of Biomedicine, Department of Microbiology and Immunology orcid

Description

Fluorescence microscopy of live yeast cells at mid-exponential growth and after 30 minutes of heat shock. The cells were grown in complete synthetic media lacking histidine and leucine with 2 % galactose as carbon source. All strains carry two plasmids; one expressing GFP-HSP104 under the control of the GAL-promoter and one expressing SSA1, SSA4 or SSA1/SSA4 chimaeras (alternatively an empty plasmid) under the control of the GPD-promoter.
The dataset was collected through fluorescence microscopy

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Data format / data structure

Still image

Data collection

Mode of collection: Biological tests

Time period(s) for data collection: 2018-06-15 — 2018-07-05

Data collector: University of Gothenburg

Source of the data: Forskningsdata: Opublicerade, Biological samples

Dataset 7

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

Suggested citation

Rebecca Andersson, Thomas Nyström. University of Gothenburg, Institute of Biomedicine, Department of Microbiology and Immunology (2020). <em>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</em>. Swedish National Data Service. Version 1. <a href="https://doi.org/10.5878/3cxh-6367">https://doi.org/10.5878/3cxh-6367</a>

Creator/Principal investigator(s)

Rebecca Andersson - University of Gothenburg, Institute of Biomedicine, Department of Microbiology and Immunology orcid

Thomas Nyström - University of Gothenburg, Institute of Biomedicine, Department of Microbiology and Immunology orcid

Description

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 Ima

... Show more..

Data format / data structure

Still image

Data collection

Mode of collection: Biological tests

Time period(s) for data collection: 2020-02-19 — 2020-02-21

Data collector: University of Gothenburg

Source of the data: Forskningsdata: Opublicerade, Biological samples

Dataset 8

Hsp70-mutant yeast strains with the misfolding marker protein guk1-7-GFP and with or without an intact HSP104-allele, imaged during mid-exponential growth

Suggested citation

Rebecca Andersson, Katarina Vielfort, Thomas Nyström. University of Gothenburg, Institute of Biomedicine, Department of Microbiology and Immunology (2020). <em>Hsp70-mutant yeast strains with the misfolding marker protein guk1-7-GFP and with or without an intact HSP104-allele, imaged during mid-exponential growth</em>. Swedish National Data Service. Version 1. <a href="https://doi.org/10.5878/d0ya-5n21">https://doi.org/10.5878/d0ya-5n21</a>

Creator/Principal investigator(s)

Katarina Vielfort - Umeå University, Institute of Molecular Biology orcid

Rebecca Andersson - University of Gothenburg, Institute of Biomedicine, Department of Microbiology and Immunology orcid

Thomas Nyström - University of Gothenburg, Institute of Biomedicine, Department of Microbiology and Immunology orcid

Description

Fluorescence microscopy of live yeast cells at mid-exponential growth. The yeast is grown in complete synthetic medium with 2 % glucose as carbon source. All strains produce the aggregate marker protein guk1-7-GFP.
The dataset was collected through fluorescence microscopy.
The image files are provided in Carl Zeiss Image format (.czi).

Data format / data structure

Still image

Data collection

Mode of collection: Biological tests

Time period(s) for data collection: 2019-04-23 — 2019-04-25

Data collector: University of Gothenburg

Source of the data: Forskningsdata: Opublicerade, Biological samples

Dataset 9

Western blots of SDS-PAGE gels with primary antibodies against Hsp70p and Pgk1p

Suggested citation

Rebecca Andersson, Thomas Nyström. University of Gothenburg, Institute of Biomedicine, Department of Microbiology and Immunology (2020). <em>Western blots of SDS-PAGE gels with primary antibodies against Hsp70p and Pgk1p</em>. Swedish National Data Service. Version 1. <a href="https://doi.org/10.5878/rq33-3f44">https://doi.org/10.5878/rq33-3f44</a>

Creator/Principal investigator(s)

Rebecca Andersson - University of Gothenburg, Institute of Biomedicine, Department of Microbiology and Immunology orcid

Thomas Nyström - University of Gothenburg, Institute of Biomedicine, Department of Microbiology and Immunology orcid

Description

WT ssa12DD and ssa12DDSSA4OE: Western blots of SDS-PAGE gels with protein extracts from yeast in mid-exponential growth. The yeast was grown in complete synthetic media with 2 % glucose as carbon source.
WT ssa12DD w chimaera plasmids: Western blots of SDS-PAGE gels with protein extracts from yeast in mid-exponential growth before and after 30 minutes of heat shock. The yeast was grown in complete synthetic media lacking leucine with 2 % glucose as carbon source.
The images are scans of wester

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Data format / data structure

Still image

Data collection

Mode of collection: Biological tests

Time period(s) for data collection: 2018-04-11 — 2018-07-24

Data collector: University of Gothenburg

Source of the data: Forskningsdata: Opublicerade, Biological samples

Dataset 10

Immunocytofluoroscense of Hsp70-mutant yeast strains with primary antibodies against Hsp42p

Suggested citation

Rebecca Andersson, Katarina Vielfort, Thomas Nyström. University of Gothenburg, Institute of Biomedicine, Department of Microbiology and Immunology (2020). <em>Immunocytofluoroscense of Hsp70-mutant yeast strains with primary antibodies against Hsp42p</em>. Swedish National Data Service. Version 1. <a href="https://doi.org/10.5878/a3kq-6708">https://doi.org/10.5878/a3kq-6708</a>

Creator/Principal investigator(s)

Katarina Vielfort - Umeå University, Institute of Molecular Biology orcid

Rebecca Andersson - University of Gothenburg, Institute of Biomedicine, Department of Microbiology and Immunology orcid

Thomas Nyström - University of Gothenburg, Institute of Biomedicine, Department of Microbiology and Immunology orcid

Description

Fluorescence microscopy of fixated yeast cells at mid-exponential growth. The samples have been stained with antibodies against Hsp42 (primary antibody: rabbit anti Hsp42, secondary antibody: goat anti rabbit Alexa Flour488) and counterstained with DAPI (datasets from 170505 and 170608) and antibodies against Actin (primary antibody: mouse anti- Act1, secondary antibody: goat anti mouse Alexa Flour 568) (dataset from 170518) or nucleoporin complexes (dataset from 170608) (primary antibody: mouse

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Data format / data structure

Still image

Data collection

Mode of collection: Biological tests

Time period(s) for data collection: 2017-05-05 — 2017-06-08

Data collector: University of Gothenburg

Source of the data: Forskningsdata: Opublicerade, Biological samples

Dataset 11

Silver stained 2D-gels of protein extracts from wild type and Hsp70-mutant yeast strains

Suggested citation

Rebecca Andersson, David Öling, Thomas Nyström. University of Gothenburg, Institute of Biomedicine, Department of Microbiology and Immunology (2020). <em>Silver stained 2D-gels of protein extracts from wild type and Hsp70-mutant yeast strains</em>. Swedish National Data Service. Version 1. <a href="https://doi.org/10.5878/9xxe-g981">https://doi.org/10.5878/9xxe-g981</a>

Creator/Principal investigator(s)

David Öling - AstraZeneca, Mölnlycke, Sweden, Discovery Biology, Discovery Sciences, Innovative Medicines and Early Development Biotech Unit orcid

Rebecca Andersson - University of Gothenburg, Institute of Biomedicine, Department of Microbiology and Immunology orcid

Thomas Nyström - University of Gothenburg, Institute of Biomedicine, Department of Microbiology and Immunology orcid

Description

Images of two dimensional protein gels with protein extracts from wild type and Hsp70 mutant yeast strains in mid-exponential growth phase. The yeast were grown in rich medium (YP) with 2 % glucose as carbon source. The images have been aligned and the background has been removed.
The dataset was collected through imaging of two dimensional protein gels.
The image files are provided in TIFF format.

Data format / data structure

Still image

Data collection

Mode of collection: Biological tests

Time period(s) for data collection: 2012-06-13 — 2013-04-10

Source of the data: Research data, Research data: Published, Biological samples

Published: 2020-12-16