title: The regulatory region of the human desmocollin 3 promoter forms a DNA four-way junction.
abstract: Adhesion between desmosomal junctions is mediated by structural proteins of the cadherin family, viz. three desmocollins (DSC) and three desmogleins (DSG). Promoter and primer extension analysis of human DSC3 showed a TATA-less sequence initiating transcription via a cluster of sites upstream of the coding region. Deletion analysis of 1 kb of the promoter showed that expression is regulated between --303 and --203 bp upstream of the start-site of translation. Tertiary structure analysis of this cis-active region (cis 1) revealed a potential DNA 4-way junction which is notably G/C-rich in sequence. PAGE analysis of this region identified four differently migrating forms of the DNA. Structure-specific cleavage of the DNA with bacteriophage T7 endonuclease I showed the slowest migrating form to be either an extended/cruciform or stacked-X 4-way junction. DNA-binding, gel retardation assays of the cis 1 region showed distinct DNA-protein complexes and by competition experiments and using purified junction DNA we show that one of these complexes bound with both sequence and structure specificity to the 4-way junction DNA.
authors.affiliation: Division of Membrane Biology, National Institute for Medical Research, Mill Hill, London NW7 1AA, United Kingdom.
authors.forename: P L
authors.initials: PL
authors.lastname: Gadhavi
authors.forename: M D
authors.initials: MD
authors.lastname: Greenwood
authors.forename: M
authors.initials: M
authors.lastname: Strom
authors.forename: I A
authors.initials: IA
authors.lastname: King
authors.forename: R S
authors.initials: RS
authors.lastname: Buxton
mesh_terms: D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D002478:Cells, Cultured; D003001:Cloning, Molecular; D004247:DNA / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D003850:Deoxyribonuclease I / Q000378:metabolism; D051187:Desmocollins; D006801:Humans; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D008562:Membrane Glycoproteins / Q000235:genetics*; D051379:Mice; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D012045:Regulatory Sequences, Nucleic Acid / Q000235:genetics*; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D017384:Sequence Deletion; D012689:Sequence Homology, Nucleic Acid; D014158:Transcription, Genetic; D014407:Tumor Cells, Cultured
chemical_list: C490356:DSC3 protein, human; D051187:Desmocollins; D008562:Membrane Glycoproteins; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; C090171:Dsc3 protein, mouse; D004247:DNA; D008156:Luciferases; D003850:Deoxyribonuclease I