HAP4, the glucose-repressed regulated subunit of the HAP transcriptional complex involved in the fermentation-respiration shift, has a functional homologue in the respiratory yeast Kluyveromyces lactis.
In Saccharomyces cerevisiae, the heteromeric HAP transcription factor is necessary for optimal growth on respiratory carbon sources. One of its components, the Hap4p protein, is necessary for transcriptional activation. The same protein is also the regulatory part of the complex in response to carbon sources, as HAP4 is strongly induced during the shift from fermentative to respiratory metabolism in S. cerevisiae. We report here the characterization of a new gene from the respiratory yeast Kluyveromyces lactis, obtained by heterologous complementation of a delta hap4 S. cerevisiae mutant strain. The deduced sequence of the protein (643 amino acids) exhibits two small domains (11 and 16 amino acids respectively) highly homologous to corresponding domains of ScHap4p, while the overall similarity is rather weak. Additional experiments were performed to confirm the functional homology of this new gene with ScHAP4, which we named KIHAP4. The importance of the small highly conserved N-terminal sequence was confirmed by in vitro mutagenesis. All the mutations that interfere with the Hap4p-Hap2/3/5 interaction were localized in it. The discovery of the same regulatory protein in two metabolically distinct yeast species raises the question of its functional significance during evolution.
Bourgarel
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Institut de Génétique et Microbiologie (URA 2225 du CNRS), Université Paris-Sud, Orsay, France.
Nguyen
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CC
Bolotin-Fukuhara
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M
D000595:Amino Acid Sequence; D023081:CCAAT-Binding Factor; D017124:Conserved Sequence; D003470:Culture Media; D003574:Cytochrome c Group / Q000378:metabolism; D045304:Cytochromes c; D005285:Fermentation; D005656:Fungal Proteins / Q000235:genetics* / Q000378:metabolism*; D015966:Gene Expression Regulation, Fungal; D005816:Genetic Complementation Test; D007716:Kluyveromyces / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism*; D007773:Lactates / Q000378:metabolism; D009154:Mutation; D010101:Oxygen Consumption; D011401:Promoter Regions, Genetic; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000254:growth & development / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors; D014158:Transcription, Genetic
D023081:CCAAT-Binding Factor; C014059:CYC1 protein, S cerevisiae; D003470:Culture Media; D003574:Cytochrome c Group; D005656:Fungal Proteins; C061437:HAP4 protein, S cerevisiae; D007773:Lactates; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors; D045304:Cytochromes c