title: Differential effect of zinc finger deletions on the binding of CTCF to the promoter of the amyloid precursor protein gene.
abstract: High levels of transcription from the amyloid precursor protein promoter are dependent on the binding of CTCF to the APBbeta core recognition sequence located between positions -82 and -93 upstream from the transcriptional start site. CTCF comprises 727 amino acids and contains 11 zinc finger motifs arranged in tandem that are flanked by 267 amino acids on the N-terminal side and 150 amino acids on the C-terminal side. Deletion of either the N- or the C-terminal regions outside of the zinc finger domain had no detrimental effect on the binding of CTCF to APBbeta. However, internal deletions of zinc fingers 5-7 completely abolished binding. The binding of full-length CTCF generated a DNase I protected domain extending from position -78 to -116, which was interrupted by a hypersensitive site at position -99. Selective deletions from the N- and C-terminal sides of the zinc finger domain showed that the N-terminal end of the zinc finger domain was aligned toward the transcriptional start site. Furthermore, deletions of zinc fingers peripheral to the essential zinc fingers 5-7 decreased the stability of the binding complex by interrupting sequence-specific interactions.
authors.affiliation: Department of Psychiatry and Behavioral Science, State University of New York at Stony Brook, Stony Brook, NY 11794-8101, USA. wquitschke@mail.psychiatry.sunysb.edu
authors.forename: W W
authors.initials: WW
authors.lastname: Quitschke
authors.forename: M J
authors.initials: MJ
authors.lastname: Taheny
authors.forename: L J
authors.initials: LJ
authors.lastname: Fochtmann
authors.forename: A A
authors.initials: AA
authors.lastname: Vostrov
mesh_terms: D000595:Amino Acid Sequence; D016564:Amyloid beta-Protein Precursor / Q000235:genetics*; D001483:Base Sequence; D001665:Binding Sites; D000076246:CCCTC-Binding Factor; D004247:DNA / Q000235:genetics / Q000378:metabolism*; D018983:DNA Footprinting; D004268:DNA-Binding Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D003850:Deoxyribonuclease I / Q000378:metabolism; D005786:Gene Expression Regulation; D006801:Humans; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic / Q000235:genetics*; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D012097:Repressor Proteins; D020218:Response Elements / Q000235:genetics; D017384:Sequence Deletion / Q000235:genetics; D013816:Thermodynamics; D014157:Transcription Factors / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D014158:Transcription, Genetic / Q000235:genetics; D016335:Zinc Fingers
chemical_list: D016564:Amyloid beta-Protein Precursor; D000076246:CCCTC-Binding Factor; C000619697:CTCF protein, human; D004268:DNA-Binding Proteins; D012097:Repressor Proteins; D014157:Transcription Factors; D004247:DNA; D003850:Deoxyribonuclease I