title: Functional study of the Hap4-like genes suggests that the key regulators of carbon metabolism HAP4 and oxidative stress response YAP1 in yeast diverged from a common ancestor.
abstract: The transcriptional regulator HAP4, induced by respiratory substrates, is involved in the balance between fermentation and respiration in S. cerevisiae. We identified putative orthologues of the Hap4 protein in all ascomycetes, based only on a conserved sixteen amino acid-long motif. In addition to this motif, some of these proteins contain a DNA-binding motif of the bZIP type, while being nonetheless globally highly divergent. The genome of the yeast Hansenula polymorpha contains two HAP4-like genes encoding the protein HpHap4-A which, like ScHap4, is devoid of a bZIP motif, and HpHap4-B which contains it. This species has been chosen for a detailed examination of their respective properties. Based mostly on global gene expression studies performed in the S. cerevisiae HAP4 disruption mutant (ScΔhap4), we show here that HpHap4-A is functionally equivalent to ScHap4, whereas HpHap4-B is not. Moreover HpHAP4-B is able to complement the H2O2 hypersensitivity of the ScYap1 deletant, YAP1 being, in S. cerevisiae, the main regulator of oxidative stress. Finally, a transcriptomic analysis performed in the ScΔyap1 strain overexpressing HpHAP4-B shows that HpHap4-B acts both on oxidative stress response and carbohydrate metabolism in a manner different from both ScYap1 and ScHap4. Deletion of these two genes in their natural host, H. polymorpha, confirms that HpHAP4-A participates in the control of the fermentation/respiration balance, while HpHAP4-B is involved in oxidative stress since its deletion leads to hypersensitivity to H2O2. These data, placed in an evolutionary context, raise new questions concerning the evolution of the HAP4 transcriptional regulation function and suggest that Yap1 and Hap4 have diverged from a unique regulatory protein in the fungal ancestor.
authors.affiliation: Institut de Génétique et Microbiologie, IFR Génome 115, Université Paris-Sud and CNRS, Orsay, France; Institute of Cell Biology, National Academy of Sciences, Lviv, Ukraine; Centre de Génétique Moléculaire, CNRS, Gif sur Yvette, France.
authors.forename: Nataliya
authors.initials: N
authors.lastname: Petryk
authors.affiliation: Institut de Génétique et Microbiologie, IFR Génome 115, Université Paris-Sud and CNRS, Orsay, France.
authors.forename: You-Fang
authors.initials: YF
authors.lastname: Zhou
authors.affiliation: Institut de Génétique et Microbiologie, IFR Génome 115, Université Paris-Sud and CNRS, Orsay, France.
authors.forename: Kateryna
authors.initials: K
authors.lastname: Sybirna
authors.affiliation: Gif/Orsay DNA MicroArray Platform, Gif sur Yvette, France; Centre de Génétique Moléculaire, CNRS, Gif sur Yvette, France.
authors.forename: Marie-Hélène
authors.initials: MH
authors.lastname: Mucchielli
authors.affiliation: Centre de Génétique Moléculaire, CNRS, Gif sur Yvette, France.
authors.forename: Bernard
authors.initials: B
authors.lastname: Guiard
authors.affiliation: Institut de Génétique et Microbiologie, IFR Génome 115, Université Paris-Sud and CNRS, Orsay, France.
authors.forename: Wei-Guo
authors.initials: WG
authors.lastname: Bao
authors.affiliation: Institute of Cell Biology, National Academy of Sciences, Lviv, Ukraine.
authors.forename: Oleh V
authors.initials: OV
authors.lastname: Stasyk
authors.affiliation: Institute of Cell Biology, National Academy of Sciences, Lviv, Ukraine; Department of Biochemistry, Ivan Franko Lviv National University, Lviv, Ukraine.
authors.forename: Olena G
authors.initials: OG
authors.lastname: Stasyk
authors.affiliation: Institute of Cell Biology, National Academy of Sciences, Lviv, Ukraine.
authors.forename: Olena S
authors.initials: OS
authors.lastname: Krasovska
authors.affiliation: Institut de Génétique et Microbiologie, IFR Génome 115, Université Paris-Sud and CNRS, Orsay, France; Gif/Orsay DNA MicroArray Platform, Gif sur Yvette, France.
authors.forename: Karine
authors.initials: K
authors.lastname: Budin
authors.affiliation: Gif/Orsay DNA MicroArray Platform, Gif sur Yvette, France; Centre de Génétique Moléculaire, CNRS, Gif sur Yvette, France.
authors.forename: Nancie
authors.initials: N
authors.lastname: Reymond
authors.affiliation: Gif/Orsay DNA MicroArray Platform, Gif sur Yvette, France.
authors.forename: Sandrine
authors.initials: S
authors.lastname: Imbeaud
authors.affiliation: Gif/Orsay DNA MicroArray Platform, Gif sur Yvette, France.
authors.forename: Sophie
authors.initials: S
authors.lastname: Coudouel
authors.affiliation: Gif/Orsay DNA MicroArray Platform, Gif sur Yvette, France; Centre de Génétique Moléculaire, CNRS, Gif sur Yvette, France.
authors.forename: Hervé
authors.initials: H
authors.lastname: Delacroix
authors.affiliation: Institute of Cell Biology, National Academy of Sciences, Lviv, Ukraine; University of Rzeszow, Rzeszow, Poland.
authors.forename: Andriy
authors.initials: A
authors.lastname: Sibirny
authors.affiliation: Institut de Génétique et Microbiologie, IFR Génome 115, Université Paris-Sud and CNRS, Orsay, France.
authors.forename: Monique
authors.initials: M
authors.lastname: Bolotin-Fukuhara
mesh_terms: D020816:Amino Acid Motifs / Q000235:genetics; D023081:CCAAT-Binding Factor / Q000235:genetics* / Q000378:metabolism; D002244:Carbon / Q000378:metabolism; D015966:Gene Expression Regulation, Fungal; D016681:Genome, Fungal; D006861:Hydrogen Peroxide / Q000737:chemistry; D010084:Oxidation-Reduction; D018384:Oxidative Stress / Q000235:genetics*; D012441:Saccharomyces cerevisiae / Q000235:genetics; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics* / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics* / Q000378:metabolism; D014158:Transcription, Genetic
chemical_list: D023081:CCAAT-Binding Factor; C061437:HAP4 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors; C086677:YAP1 protein, S cerevisiae; D002244:Carbon; D006861:Hydrogen Peroxide