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Study of the yeast cytosolic Hsp70-system in protein homeostasis and life span regulation - Immunocytofluoroscense of Hsp70-mutant yeast strains with primary antibodies against Hsp42p

SND-ID: 2020-36-10. Version: 1. DOI: https://doi.org/10.5878/a3kq-6708

Associated documentation

Citation

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

Research principal

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

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.

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 Alex

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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 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 anti- Nsp1, secondary antibody: goat anti mouse Alexa Flour 568).
The dataset was collected through fluorescence microscopy.
The image files are provided in Carl Zeiss Image format (.czi). Show less..

Data contains personal data

No

Language

Method and outcome

Unit of analysis

Population

Saccharomyces cerevisiae (Baker's yeast)

Study design

Experimental study

Preclinical study

Sampling procedure

Time period(s) investigated

2012 – 2020

Data format / data structure

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: Research data: Unpublished, Biological samples, Research data
Geographic coverage
Administrative information

Responsible department/unit

Institute of Biomedicine, Department of Microbiology and Immunology

Funding

  • Funding agency: Knut and Alice Wallenberg Foundation rorId
Topic and keywords

Research area

Biochemistry and molecular biology (Standard för svensk indelning av forskningsämnen 2011)

Cell biology (Standard för svensk indelning av forskningsämnen 2011)

Microbiology (Standard för svensk indelning av forskningsämnen 2011)

Cell and molecular biology (Standard för svensk indelning av forskningsämnen 2011)

Keywords

Molecular chaperones, Hsp70 heat-shock proteins, Proteostasis deficiencies

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, et al. (2021) Differential role of cytosolic Hsp70s in longevity assurance and protein quality control. PLoS Genet 17(1): e1008951.
DOI: https://doi.org/10.1371/journal.pgen.1008951

If you have published anything based on these data, please notify us with a reference to your publication(s). If you are responsible for the catalogue entry, you can update the metadata/data description in DORIS.

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License

CC BY 4.0

Versions

Version 1. 2020-12-16

Version 1: 2020-12-16

DOI: https://doi.org/10.5878/a3kq-6708

Contact for questions about the data

Related research data in SND's catalogue

Study of the yeast cytosolic Hsp70-system in protein homeostasis and life span regulation - The misfolding protein marker gus1-3-GFP and the metacaspase Mca1-GFP during mid-exponential growth in yeast Hsp70-mutant yeast strains

Study of the yeast cytosolic Hsp70-system in protein homeostasis and life span regulation - The misfolding protein marker guk1-7-GFP before, during and after in recovery from heat shock stress in Hsp70-mutant yeast strains

Study of the yeast cytosolic Hsp70-system in protein homeostasis and life span regulation - Intracellular colocalisation of the chaperone Ssa4-GFP and the metacaspase Mca1-RFP before and after heat stress in a Hsp70-mutant yeast cell strain

Study of the yeast cytosolic Hsp70-system in protein homeostasis and life span regulation - The molecular chaperone GFP-Hsp104 before and after heat stress in Hsp70-mutant yeast strains

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

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

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

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

Study of the yeast cytosolic Hsp70-system in protein homeostasis and life span regulation - Western blots of SDS-PAGE gels with primary antibodies against Hsp70p and Pgk1p

Study of the yeast cytosolic Hsp70-system in protein homeostasis and life span regulation - Silver stained 2D-gels of protein extracts from wild type and Hsp70-mutant yeast strains

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Published: 2020-12-16
Last updated: 2021-02-11