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Depletion of yeast PDK1 orthologs triggers a stress-like transcriptional response

BACKGROUND: Pkh proteins are the PDK1 orthologs in S. cerevisiae. They have redundant and essential activity and are responsible for the phosphorylation of several members of the AGC family of protein kinases. Pkh proteins have been involved in several cellular functions, including cell wall integri...

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Autores principales: Pastor-Flores, Daniel, Ferrer-Dalmau, Jofre, Bahí, Anna, Boleda, Martí, Biondi, Ricardo M., Casamayor, Antonio
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4578605/
https://www.ncbi.nlm.nih.gov/pubmed/26391581
http://dx.doi.org/10.1186/s12864-015-1903-8
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author Pastor-Flores, Daniel
Ferrer-Dalmau, Jofre
Bahí, Anna
Boleda, Martí
Biondi, Ricardo M.
Casamayor, Antonio
author_facet Pastor-Flores, Daniel
Ferrer-Dalmau, Jofre
Bahí, Anna
Boleda, Martí
Biondi, Ricardo M.
Casamayor, Antonio
author_sort Pastor-Flores, Daniel
collection PubMed
description BACKGROUND: Pkh proteins are the PDK1 orthologs in S. cerevisiae. They have redundant and essential activity and are responsible for the phosphorylation of several members of the AGC family of protein kinases. Pkh proteins have been involved in several cellular functions, including cell wall integrity and endocytosis. However the global expression changes caused by their depletion are still unknown. RESULTS: A doxycycline-repressible tetO(7) promoter driving the expression of PKH2 in cells carrying deletions of the PKH1 and PKH3 genes allowed us to progressively deplete cells from Pkh proteins when treated with doxycycline. Global gene expression analysis indicate that depletion of Pkh results in the up-regulation of genes involved in the accumulation of glycogen and also of those related to stress responses. Moreover, genes involved in the ion transport were quickly down-regulated when the levels of Pkh decreased. The reduction in the mRNA levels required for protein translation, however, was only observed after longer doxycycline treatment (24 h). We uncovered that Pkh is important for the proper transcriptional response to heat shock, and is mostly required for the effects driven by the transcription factors Hsf1 and Msn2/Msn4, but is not required for down-regulation of the mRNA coding for ribosomal proteins. CONCLUSIONS: By using the tetO(7) promoter we elucidated for the first time the transcriptomic changes directly or indirectly caused by progressive depletion of Pkh. Furthermore, this system enabled the characterization of the transcriptional response triggered by heat shock in wild-type and Pkh-depleted cells, showing that about 40 % of the observed expression changes were, to some degree, dependent on Pkh. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12864-015-1903-8) contains supplementary material, which is available to authorized users.
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spelling pubmed-45786052015-09-23 Depletion of yeast PDK1 orthologs triggers a stress-like transcriptional response Pastor-Flores, Daniel Ferrer-Dalmau, Jofre Bahí, Anna Boleda, Martí Biondi, Ricardo M. Casamayor, Antonio BMC Genomics Research Article BACKGROUND: Pkh proteins are the PDK1 orthologs in S. cerevisiae. They have redundant and essential activity and are responsible for the phosphorylation of several members of the AGC family of protein kinases. Pkh proteins have been involved in several cellular functions, including cell wall integrity and endocytosis. However the global expression changes caused by their depletion are still unknown. RESULTS: A doxycycline-repressible tetO(7) promoter driving the expression of PKH2 in cells carrying deletions of the PKH1 and PKH3 genes allowed us to progressively deplete cells from Pkh proteins when treated with doxycycline. Global gene expression analysis indicate that depletion of Pkh results in the up-regulation of genes involved in the accumulation of glycogen and also of those related to stress responses. Moreover, genes involved in the ion transport were quickly down-regulated when the levels of Pkh decreased. The reduction in the mRNA levels required for protein translation, however, was only observed after longer doxycycline treatment (24 h). We uncovered that Pkh is important for the proper transcriptional response to heat shock, and is mostly required for the effects driven by the transcription factors Hsf1 and Msn2/Msn4, but is not required for down-regulation of the mRNA coding for ribosomal proteins. CONCLUSIONS: By using the tetO(7) promoter we elucidated for the first time the transcriptomic changes directly or indirectly caused by progressive depletion of Pkh. Furthermore, this system enabled the characterization of the transcriptional response triggered by heat shock in wild-type and Pkh-depleted cells, showing that about 40 % of the observed expression changes were, to some degree, dependent on Pkh. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12864-015-1903-8) contains supplementary material, which is available to authorized users. BioMed Central 2015-09-21 /pmc/articles/PMC4578605/ /pubmed/26391581 http://dx.doi.org/10.1186/s12864-015-1903-8 Text en © Pastor-Flores et al. 2015 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research Article
Pastor-Flores, Daniel
Ferrer-Dalmau, Jofre
Bahí, Anna
Boleda, Martí
Biondi, Ricardo M.
Casamayor, Antonio
Depletion of yeast PDK1 orthologs triggers a stress-like transcriptional response
title Depletion of yeast PDK1 orthologs triggers a stress-like transcriptional response
title_full Depletion of yeast PDK1 orthologs triggers a stress-like transcriptional response
title_fullStr Depletion of yeast PDK1 orthologs triggers a stress-like transcriptional response
title_full_unstemmed Depletion of yeast PDK1 orthologs triggers a stress-like transcriptional response
title_short Depletion of yeast PDK1 orthologs triggers a stress-like transcriptional response
title_sort depletion of yeast pdk1 orthologs triggers a stress-like transcriptional response
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4578605/
https://www.ncbi.nlm.nih.gov/pubmed/26391581
http://dx.doi.org/10.1186/s12864-015-1903-8
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