The universally conserved P-loop ATPase Ola1 is implicated in various cellular stress response pathways, as well as in cancer and tumor progression. However, Ola1p functions are divergent between species, and the involved mechanisms are only poorly understood. Here, we studied the role of Ola1p in the heat shock response of the yeast Saccharomyces cerevisiae using a combination of quantitative and pulse labeling-based proteomics approaches, in~vitro studies, and cell-based assays. Our data show that when heat stress is applied to cells lacking Ola1p, the expression of stress-protective proteins is enhanced. During heat stress Ola1p associates with detergent-resistant protein aggregates and rapidly forms assemblies that localize to stress granules. The assembly of Ola1p was also observed in~vitro using purified protein and conditions, which resembled those in living cells. We show that loss of Ola1p results in increased protein ubiquitination of detergent-insoluble aggregates recovered from heat-shocked cells. When cells lacking Ola1p were subsequently relieved from heat stress, reinitiation of translation was delayed, whereas, at the same time, de novo synthesis of central factors required for protein refolding and the clearance of aggregates was enhanced when compared with wild-type cells. The combined data suggest that upon acute heat stress, Ola1p is involved in the stabilization of misfolded proteins, which become sequestered in cytoplasmic stress granules. This function of Ola1p enables cells to resume translation in a timely manner as soon as heat stress is relieved.
%0 Journal Article
%1 dannenmaierQuantitativeProteomicsIdentifies2021a
%A Dannenmaier, Stefan
%A Desroches Altamirano, Christine
%A Schüler, Lisa
%A Zhang, Ying
%A Hummel, Johannes
%A Milanov, Martin
%A Oeljeklaus, Silke
%A Koch, Hans-Georg
%A Rospert, Sabine
%A Alberti, Simon
%A Warscheid, Bettina
%C United States
%D 2021
%J The Journal of biological chemistry
%K *Gene 1,heat Biosynthesis,Adenosine Expression Fungal,*Heat-Shock Proteins/genetics/*metabolism,Saccharomyces Regulation Response,*Protein Triphosphatases/genetics/*metabolism,heat aggregation,protein cerevisiae cerevisiae/genetics/*metabolism,stress factor granule,to_read,translation initiation misfolding,protein protein protein,protein self-assembly,protein shock synthesis,proteomics,Saccharomyces
%N 5
%P 101050
%R 10.1016/j.jbc.2021.101050
%T Quantitative Proteomics Identifies the Universally Conserved ATPase Ola1p as a Positive Regulator of Heat Shock Response in Saccharomyces Cerevisiae.
%V 297
%X The universally conserved P-loop ATPase Ola1 is implicated in various cellular stress response pathways, as well as in cancer and tumor progression. However, Ola1p functions are divergent between species, and the involved mechanisms are only poorly understood. Here, we studied the role of Ola1p in the heat shock response of the yeast Saccharomyces cerevisiae using a combination of quantitative and pulse labeling-based proteomics approaches, in~vitro studies, and cell-based assays. Our data show that when heat stress is applied to cells lacking Ola1p, the expression of stress-protective proteins is enhanced. During heat stress Ola1p associates with detergent-resistant protein aggregates and rapidly forms assemblies that localize to stress granules. The assembly of Ola1p was also observed in~vitro using purified protein and conditions, which resembled those in living cells. We show that loss of Ola1p results in increased protein ubiquitination of detergent-insoluble aggregates recovered from heat-shocked cells. When cells lacking Ola1p were subsequently relieved from heat stress, reinitiation of translation was delayed, whereas, at the same time, de novo synthesis of central factors required for protein refolding and the clearance of aggregates was enhanced when compared with wild-type cells. The combined data suggest that upon acute heat stress, Ola1p is involved in the stabilization of misfolded proteins, which become sequestered in cytoplasmic stress granules. This function of Ola1p enables cells to resume translation in a timely manner as soon as heat stress is relieved.
@article{dannenmaierQuantitativeProteomicsIdentifies2021a,
abstract = {The universally conserved P-loop ATPase Ola1 is implicated in various cellular stress response pathways, as well as in cancer and tumor progression. However, Ola1p functions are divergent between species, and the involved mechanisms are only poorly understood. Here, we studied the role of Ola1p in the heat shock response of the yeast Saccharomyces cerevisiae using a combination of quantitative and pulse labeling-based proteomics approaches, in~vitro studies, and cell-based assays. Our data show that when heat stress is applied to cells lacking Ola1p, the expression of stress-protective proteins is enhanced. During heat stress Ola1p associates with detergent-resistant protein aggregates and rapidly forms assemblies that localize to stress granules. The assembly of Ola1p was also observed in~vitro using purified protein and conditions, which resembled those in living cells. We show that loss of Ola1p results in increased protein ubiquitination of detergent-insoluble aggregates recovered from heat-shocked cells. When cells lacking Ola1p were subsequently relieved from heat stress, reinitiation of translation was delayed, whereas, at the same time, de novo synthesis of central factors required for protein refolding and the clearance of aggregates was enhanced when compared with wild-type cells. The combined data suggest that upon acute heat stress, Ola1p is involved in the stabilization of misfolded proteins, which become sequestered in cytoplasmic stress granules. This function of Ola1p enables cells to resume translation in a timely manner as soon as heat stress is relieved.},
added-at = {2024-05-17T13:01:35.000+0200},
address = {United States},
author = {Dannenmaier, Stefan and Desroches Altamirano, Christine and Sch{\"u}ler, Lisa and Zhang, Ying and Hummel, Johannes and Milanov, Martin and Oeljeklaus, Silke and Koch, Hans-Georg and Rospert, Sabine and Alberti, Simon and Warscheid, Bettina},
biburl = {https://www.bibsonomy.org/bibtex/2de571919471e23793464861778ce1b45/warscheidlab},
copyright = {Copyright {\copyright} 2021 The Authors. Published by Elsevier Inc. All rights reserved.},
doi = {10.1016/j.jbc.2021.101050},
interhash = {c79fdd882c0cd501f2d49af6cd29c6a1},
intrahash = {de571919471e23793464861778ce1b45},
issn = {1083-351X 0021-9258},
journal = {The Journal of biological chemistry},
keywords = {*Gene 1,heat Biosynthesis,Adenosine Expression Fungal,*Heat-Shock Proteins/genetics/*metabolism,Saccharomyces Regulation Response,*Protein Triphosphatases/genetics/*metabolism,heat aggregation,protein cerevisiae cerevisiae/genetics/*metabolism,stress factor granule,to_read,translation initiation misfolding,protein protein protein,protein self-assembly,protein shock synthesis,proteomics,Saccharomyces},
langid = {english},
month = nov,
number = 5,
pages = 101050,
pmcid = {PMC8531669},
pmid = {34571008},
timestamp = {2024-05-17T13:01:35.000+0200},
title = {Quantitative Proteomics Identifies the Universally Conserved {{ATPase Ola1p}} as a Positive Regulator of Heat Shock Response in {{Saccharomyces}} Cerevisiae.},
volume = 297,
year = 2021
}