title: Distinct class of DNA-binding domains is exemplified by a master regulator of phenotypic switching in Candida albicans.
abstract: Among the most important classes of regulatory proteins are the sequence-specific DNA-binding proteins that control transcription through the occupancy of discrete DNA sequences within genomes. Currently, this class of proteins encompasses at least 37 distinct structural superfamilies and more than 100 distinct structural motifs. In this paper, we examine the transcriptional regulator Wor1, a master regulator of white-opaque switching in the human fungal pathogen Candida albicans. As assessed by a variety of algorithms, this protein has no sequence or structural similarity to any known DNA-binding protein. It is, however, conserved across the vast fungal lineage, with a 300aa region of sequence conservation. Here, we show that this 300aa region of Wor1 exhibits sequence-specific DNA binding and therefore represents a new superfamily of DNA-binding proteins. We identify the 14-nucleotide-pair DNA sequence recognized by Wor1, characterize the site through mutational analysis, and demonstrate that this sequence is sufficient for the Wor1-dependent activation of transcription in vivo. Within the 300aa DNA-binding conserved region, which we have termed the WOPR box, are two domains (WOPRa and WOPRb), dissimilar to each other but especially well-conserved across the fungal lineage. We show that the WOPR box binds DNA as a monomer and that neither domain, when expressed and purified separately, exhibits sequence-specific binding. DNA binding is restored, however, when the two isolated domains are added together. These results indicate that the WOPR family of DNA-binding proteins involves an unusual coupling between two dissimilar, covalently linked domains.
authors.affiliation: Department of Biochemistry and Biophysics, 600 16th Street, University of California, San Francisco, CA 94158, USA.
authors.forename: Matthew B
authors.initials: MB
authors.lastname: Lohse
authors.forename: Rebecca E
authors.initials: RE
authors.lastname: Zordan
authors.forename: Christopher W
authors.initials: CW
authors.lastname: Cain
authors.forename: Alexander D
authors.initials: AD
authors.lastname: Johnson
mesh_terms: D002176:Candida albicans / Q000737:chemistry* / Q000235:genetics; D017124:Conserved Sequence; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000235:genetics*; D005656:Fungal Proteins; D015966:Gene Expression Regulation, Fungal; D006801:Humans; D010641:Phenotype; D014157:Transcription Factors / Q000737:chemistry*; D014158:Transcription, Genetic
chemical_list: D004268:DNA-Binding Proteins; D005656:Fungal Proteins; D014157:Transcription Factors