title: The C-terminal region of the Hot1 transcription factor binds GGGACAAA-related sequences in the promoter of its target genes.
abstract: Response to hyperosmotic stress in the yeast Saccharomyces cerevisiae involves the participation of the general stress response mediated by Msn2/4 transcription factors and the HOG pathway. One of the transcription factors activated through this pathway is Hot1, which contributes to the control of the expression of several genes involved in glycerol synthesis and flux, or in other functions related to adaptation to adverse conditions. This work provides new data about the interaction mechanism of this transcription factor with DNA. By means of one-hybrid and electrophoretic mobility assays, we demonstrate that the C-terminal region, which corresponds to amino acids 610-719, is the DNA-binding domain of Hot1. We also describe how this domain recognizes sequence 5'-GGGACAAA-3' located in the promoter of gene STL1. The bioinformatics analysis carried out in this work allowed the identification of identical or similar sequences (with up to two mismatches) in the promoter of other Hot1 targets, where central element GGACA was quite conserved among them. Finally, we found that small variations in the sequence recognized by Hot1 may influence its ability to recognize its targets in vivo.
authors.affiliation: Departament de Bioquímica i Biologia Molecular, Facultat de Ciències Biològiques, Universitat de València, Burjassot, Valencia, Spain.
authors.forename: Mercè
authors.initials: M
authors.lastname: Gomar-Alba
authors.affiliation: Genomics and Stress Laboratory, Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, Oeiras, Portugal.
authors.forename: Catarina
authors.initials: C
authors.lastname: Amaral
authors.affiliation: Joint Unit of Research in Genomics and Health, Fundación para el Fomento de la Investigación Sanitaria y Biomédica de la Comunidad Valenciana (FISABIO)-Salud Pública, Valencia, Spain.
authors.forename: Alejandro
authors.initials: A
authors.lastname: Artacho
authors.affiliation: Joint Unit of Research in Genomics and Health, Fundación para el Fomento de la Investigación Sanitaria y Biomédica de la Comunidad Valenciana (FISABIO)-Salud Pública, Valencia, Spain; Centro de Investigación en Red en Epidemiología y Salud Pública (CIBEResp), Madrid, Spain.
authors.forename: Giuseppe
authors.initials: G
authors.lastname: D'Auria
authors.affiliation: Genomics and Stress Laboratory, Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, Oeiras, Portugal.
authors.forename: Catarina
authors.initials: C
authors.lastname: Pimentel
authors.affiliation: Genomics and Stress Laboratory, Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, Oeiras, Portugal.
authors.forename: Claudina
authors.initials: C
authors.lastname: Rodrigues-Pousada
authors.affiliation: Departament de Bioquímica i Biologia Molecular, Facultat de Ciències Biològiques, Universitat de València, Burjassot, Valencia, Spain. Electronic address: m.del.olmo@uv.es.
authors.forename: Marcel
authors.initials: M
authors.lastname: lí del Olmo
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D001665:Binding Sites; D003198:Computer Simulation; D017124:Conserved Sequence; D004271:DNA, Fungal / Q000235:genetics / Q000378:metabolism*; D015966:Gene Expression Regulation, Fungal; D005800:Genes, Fungal; D026901:Membrane Transport Proteins / Q000235:genetics*; D008969:Molecular Sequence Data; D009154:Mutation; D064587:Osmoregulation / Q000235:genetics; D011401:Promoter Regions, Genetic / Q000235:genetics*; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics* / Q000378:metabolism*; D016415:Sequence Alignment; D017384:Sequence Deletion; D017386:Sequence Homology, Amino Acid; D014157:Transcription Factors / Q000378:metabolism*
chemical_list: D004271:DNA, Fungal; C120535:HOT1 protein, S cerevisiae; D026901:Membrane Transport Proteins; D011993:Recombinant Fusion Proteins; C470555:STL1 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors
keywords: DNA binding domain; Hot1; Osmotic stress; Saccharomyces cerevisiae; Sequence recognition