title: DOF-binding sites additively contribute to guard cell-specificity of AtMYB60 promoter.
abstract: BACKGROUND
  We previously demonstrated that the Arabidopsis thaliana AtMYB60 protein is an R2R3MYB transcription factor required for stomatal opening. AtMYB60 is specifically expressed in guard cells and down-regulated at the transcriptional levels by the phytohormone ABA.
  
  
  RESULTS
  To investigate the molecular mechanisms governing AtMYB60 expression, its promoter was dissected through deletion and mutagenesis analyses. By studying different versions of AtMYB60 promoter::GUS reporter fusions in transgenic plants we were able to demonstrate a modular organization for the AtMYB60 promoter. Particularly we defined: a minimal promoter sufficient to confer guard cell-specific activity to the reporter gene; the distinct roles of different DOF-binding sites organised in a cluster in the minimal promoter in determining guard cell-specific expression; the promoter regions responsible for the enhancement of activity in guard cells; a promoter region responsible for the negative transcriptional regulation by ABA. Moreover from the analysis of single and multiple mutants we could rule out the involvement of a group of DOF proteins, known as CDFs, already characterised for their involvement in flowering time, in the regulation of AtMYB60 expression.
  
  
  CONCLUSIONS
  These findings shed light on the regulation of gene expression in guard cells and provide new promoter modules as useful tools for manipulating gene expression in guard cells, both for physiological studies and future biotechnological applications.
authors.affiliation: Dipartimento di Scienze Biomolecolari e Biotecnologie, Università degli Studi di Milano, Milano, Italy. cominelli@ibba.cnr.it
authors.forename: Eleonora
authors.initials: E
authors.lastname: Cominelli
authors.forename: Massimo
authors.initials: M
authors.lastname: Galbiati
authors.forename: Alessandra
authors.initials: A
authors.lastname: Albertini
authors.forename: Fabio
authors.initials: F
authors.lastname: Fornara
authors.forename: Lucio
authors.initials: L
authors.lastname: Conti
authors.forename: George
authors.initials: G
authors.lastname: Coupland
authors.forename: Chiara
authors.initials: C
authors.lastname: Tonelli
mesh_terms: D000040:Abscisic Acid / Q000502:physiology; D017360:Arabidopsis / Q000235:genetics* / Q000502:physiology; D029681:Arabidopsis Proteins / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence; D001665:Binding Sites; D018506:Gene Expression Regulation, Plant; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D054046:Plant Stomata / Q000502:physiology*; D030821:Plants, Genetically Modified / Q000235:genetics / Q000502:physiology; D011401:Promoter Regions, Genetic; D014157:Transcription Factors / Q000235:genetics* / Q000378:metabolism
chemical_list: D029681:Arabidopsis Proteins; C504083:MYB60 protein, Arabidopsis; D014157:Transcription Factors; D000040:Abscisic Acid
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