title: Depletion of yeast PDK1 orthologs triggers a stress-like transcriptional response.
abstract: 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 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 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-type and Pkh-depleted cells, showing that about 40 % of the observed expression changes were, to some degree, dependent on Pkh.
authors.affiliation: Research Group PhosphoSites, Medizinische Klinik I, Universitätsklinikum Frankfurt, Theodor-Stern-Kai 7, 60590, Frankfurt, Germany. d.pastor-flores@dkfz.de.|Present address: Division of Redox Regulation, German Cancer Research Center, DKFZ-ZMBH Alliance, Im Neuenheimer Feld 280, 69120, Heidelberg, Germany. d.pastor-flores@dkfz.de.
authors.forename: Daniel
authors.initials: D
authors.lastname: Pastor-Flores
authors.affiliation: Departament de Bioquímica i Biologia Molecular, Facultat de Veterinària, Universitat Autònoma de Barcelona, Cerdanyola 08193, Barcelona, Spain. jofre.ferrerdalmau@gmail.com.|Institut de Biotecnologia i Biomedicina, Universitat Autònoma de Barcelona, Cerdanyola 08193, Barcelona, Spain. jofre.ferrerdalmau@gmail.com.
authors.forename: Jofre
authors.initials: J
authors.lastname: Ferrer-Dalmau
authors.affiliation: Departament de Bioquímica i Biologia Molecular, Facultat de Veterinària, Universitat Autònoma de Barcelona, Cerdanyola 08193, Barcelona, Spain. anbasala@gmail.com.|Institut de Biotecnologia i Biomedicina, Universitat Autònoma de Barcelona, Cerdanyola 08193, Barcelona, Spain. anbasala@gmail.com.
authors.forename: Anna
authors.initials: A
authors.lastname: Bahí
authors.affiliation: Departament de Bioquímica i Biologia Molecular, Facultat de Veterinària, Universitat Autònoma de Barcelona, Cerdanyola 08193, Barcelona, Spain. marti.boleda@gmail.com.|Laboratoire d'Ecologie Alpine (LECA), UMR 5553, CNRS-Université Joseph Fourie, BP 53, 38041, Grenoble, France. marti.boleda@gmail.com.
authors.forename: Martí
authors.initials: M
authors.lastname: Boleda
authors.affiliation: Research Group PhosphoSites, Medizinische Klinik I, Universitätsklinikum Frankfurt, Theodor-Stern-Kai 7, 60590, Frankfurt, Germany. biondi@med.uni-frankfurt.de.
authors.forename: Ricardo M
authors.initials: RM
authors.lastname: Biondi
authors.affiliation: Departament de Bioquímica i Biologia Molecular, Facultat de Veterinària, Universitat Autònoma de Barcelona, Cerdanyola 08193, Barcelona, Spain. antonio.casamayor@uab.cat.|Institut de Biotecnologia i Biomedicina, Universitat Autònoma de Barcelona, Cerdanyola 08193, Barcelona, Spain. antonio.casamayor@uab.cat.
authors.forename: Antonio
authors.initials: A
authors.lastname: Casamayor
mesh_terms: D064413:3-Phosphoinositide-Dependent Protein Kinases / Q000235:genetics* / Q000378:metabolism; D016000:Cluster Analysis; D005285:Fermentation; D020869:Gene Expression Profiling; D015966:Gene Expression Regulation, Fungal; D005804:Genes, Lethal; D006003:Glycogen / Q000378:metabolism; D018869:Heat-Shock Response / Q000235:genetics; D007477:Ions; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism; D013312:Stress, Physiological / Q000235:genetics*; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D014158:Transcription, Genetic
chemical_list: D007477:Ions; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors; D006003:Glycogen; D064413:3-Phosphoinositide-Dependent Protein Kinases