THAP proteins target specific DNA sites through bipartite recognition of adjacent major and minor grooves

Nat Struct Mol Biol. 2010 Jan;17(1):117-23. doi: 10.1038/nsmb.1742. Epub 2009 Dec 13.

Abstract

THAP-family C(2)CH zinc-coordinating DNA-binding proteins function in diverse eukaryotic cellular processes, such as transposition, transcriptional repression, stem-cell pluripotency, angiogenesis and neurological function. To determine the molecular basis for sequence-specific DNA recognition by THAP proteins, we solved the crystal structure of the Drosophila melanogaster P element transposase THAP domain (DmTHAP) in complex with a natural 10-base-pair site. In contrast to C(2)H(2) zinc fingers, DmTHAP docks a conserved beta-sheet into the major groove and a basic C-terminal loop into the adjacent minor groove. We confirmed specific protein-DNA interactions by mutagenesis and DNA-binding assays. Sequence analysis of natural and in vitro-selected binding sites suggests that several THAPs (DmTHAP and human THAP1 and THAP9) recognize a bipartite TXXGGGX(A/T) consensus motif; homology suggests THAP proteins bind DNA through a bipartite interaction. These findings reveal the conserved mechanisms by which THAP-family proteins engage specific chromosomal target elements.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Chromatography, Gel
  • Crystallization
  • DNA / metabolism*
  • DNA Primers / genetics
  • DNA-Binding Proteins / chemistry*
  • DNA-Binding Proteins / genetics
  • DNA-Binding Proteins / metabolism
  • Drosophila melanogaster / enzymology*
  • Electrophoretic Mobility Shift Assay
  • Models, Molecular*
  • Molecular Sequence Data
  • Mutagenesis
  • Protein Binding
  • Protein Conformation*
  • Protein Structure, Tertiary / genetics*
  • Sequence Analysis, DNA
  • Transposases / chemistry*
  • Transposases / genetics
  • Transposases / metabolism

Substances

  • DNA Primers
  • DNA-Binding Proteins
  • DNA
  • THAP9 protein, human
  • Transposases

Associated data

  • PDB/3KDE