Depletion of yeast PDK1 orthologs triggers a stress-like transcriptional response
Pastor-Flores, Daniel
Ferrer-Dalmau, Jofre (Universitat Autònoma de Barcelona. Institut de Biotecnologia i de Biomedicina "Vicent Villar Palasí")
Bahí, Anna (Universitat Autònoma de Barcelona. Departament de Bioquímica i de Biologia Molecular)
Boleda, Martí (Universitat Autònoma de Barcelona. Departament de Bioquímica i de Biologia Molecular)
Biondi, Ricardo M.
Casamayor Gracia, Antonio (Universitat Autònoma de Barcelona. Departament de Bioquímica i de Biologia Molecular)

Data:	2015
Resum:	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 tetO7 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 long er 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 Hsf1andMsn2/Msn4,butisnotrequiredfordown-reg ulation of the mRNA coding for ribosomal proteins. Conclusions: By using the tetO7 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-typeandPkh-depletedcells,showingthatabout40%ofthe observed expression changes were, to some degree, dependent on Pkh.
Drets:	Aquest document està subjecte a una llicència d'ús Creative Commons. Es permet la reproducció total o parcial, la distribució, la comunicació pública de l'obra i la creació d'obres derivades, fins i tot amb finalitats comercials, sempre i quan es reconegui l'autoria de l'obra original.
Llengua:	Anglès
Document:	Article ; recerca ; Versió publicada
Matèria:	Yeast ; Pkh protein kinases ; DNA Microarray ; Doxycycline-repressive promoter ; Heat shock
Publicat a:	BMC genomics, Vol. 16 (2015) , p. 719, ISSN 1471-2164

DOI: 10.1186/s12864-015-1903-8
PMID: 26391581

21 p, 2.1 MB

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