title: The multi-zinc finger protein ZNF217 contacts DNA through a two-finger domain.
abstract: Classical C2H2 zinc finger proteins are among the most abundant transcription factors found in eukaryotes, and the mechanisms through which they recognize their target genes have been extensively investigated. In general, a tandem array of three fingers separated by characteristic TGERP links is required for sequence-specific DNA recognition. Nevertheless, a significant number of zinc finger proteins do not contain a hallmark three-finger array of this type, raising the question of whether and how they contact DNA. We have examined the multi-finger protein ZNF217, which contains eight classical zinc fingers. ZNF217 is implicated as an oncogene and in repressing the E-cadherin gene. We show that two of its zinc fingers, 6 and 7, can mediate contacts with DNA. We examine its putative recognition site in the E-cadherin promoter and demonstrate that this is a suboptimal site. NMR analysis and mutagenesis is used to define the DNA binding surface of ZNF217, and we examine the specificity of the DNA binding activity using fluorescence anisotropy titrations. Finally, sequence analysis reveals that a variety of multi-finger proteins also contain two-finger units, and our data support the idea that these may constitute a distinct subclass of DNA recognition motif.
authors: School of Molecular Bioscience, University of Sydney, New South Wales 2006, Australia.
authors: Noelia
authors: N
authors: Nunez
authors: School of Molecular Bioscience, University of Sydney, New South Wales 2006, Australia.
authors: Molly M K
authors: MMK
authors: Clifton
authors: School of Molecular Bioscience, University of Sydney, New South Wales 2006, Australia; School of Biotechnology and Biomolecular Sciences, University of New South Wales, New South Wales 2052, Australia.
authors: Alister P W
authors: APW
authors: Funnell
authors: School of Biotechnology and Biomolecular Sciences, University of New South Wales, New South Wales 2052, Australia.
authors: Crisbel
authors: C
authors: Artuz
authors: School of Molecular Bioscience, University of Sydney, New South Wales 2006, Australia.
authors: Samantha
authors: S
authors: Hallal
authors: School of Molecular Bioscience, University of Sydney, New South Wales 2006, Australia.
authors: Kate G R
authors: KGR
authors: Quinlan
authors: School of Molecular Bioscience, University of Sydney, New South Wales 2006, Australia.
authors: Josep
authors: J
authors: Font
authors: School of Molecular Bioscience, University of Sydney, New South Wales 2006, Australia.
authors: Marylène
authors: M
authors: Vandevenne
authors: School of Molecular Bioscience, University of Sydney, New South Wales 2006, Australia.
authors: Surya
authors: S
authors: Setiyaputra
authors: School of Molecular Bioscience, University of Sydney, New South Wales 2006, Australia; School of Biotechnology and Biomolecular Sciences, University of New South Wales, New South Wales 2052, Australia.
authors: Richard C M
authors: RCM
authors: Pearson
authors: School of Molecular Bioscience, University of Sydney, New South Wales 2006, Australia.
authors: Joel P
authors: JP
authors: Mackay
authors: School of Molecular Bioscience, University of Sydney, New South Wales 2006, Australia; School of Biotechnology and Biomolecular Sciences, University of New South Wales, New South Wales 2052, Australia. Electronic address: m.crossley@unsw.edu.au.
authors: Merlin
authors: M
authors: Crossley
mesh_terms: D020816:Amino Acid Motifs; D001665:Binding Sites; D002467:Cell Nucleus / Q000378:metabolism; D004247:DNA / Q000737:chemistry*; D005786:Gene Expression Regulation; D057809:HEK293 Cells; D006801:Humans; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D008958:Models, Molecular; D011485:Protein Binding; D025941:Protein Interaction Mapping; D017434:Protein Structure, Tertiary; D015534:Trans-Activators / Q000737:chemistry / Q000502:physiology*; D014158:Transcription, Genetic; D016335:Zinc Fingers
chemical_list: D015534:Trans-Activators; C113698:ZNF217 protein, human; D004247:DNA