Differences and similarities in DNA-binding preferences of MyoD and E2A protein complexes revealed by binding site selection

Science. 1990 Nov 23;250(4984):1104-10. doi: 10.1126/science.2174572.

Abstract

A technique was developed for studying protein-DNA recognition that can be applied to any purified protein, partially purified protein, or cloned gene. From oligonucleotides in which particular positions are of random sequence, that subset to which a given protein binds is amplified by the polymerase chain reaction and sequenced as a pool. These selected and amplified binding site (SAAB) "imprints" provide a characteristic set of preferred sequences for protein binding. With this technique, it was shown that homo- and heterooligomers of the helix-loop-helix proteins MyoD and E2A recognize a common consensus sequence, CA--TG, but otherwise bind to flanking and internal positions with different sequence preferences that suggest half-site recognition. These findings suggest that different combinations of dimeric proteins can have different binding sequence preferences.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • Binding Sites
  • DNA / metabolism*
  • DNA-Binding Proteins / metabolism*
  • Glutathione Transferase
  • Macromolecular Substances
  • Molecular Sequence Data
  • Muscle Proteins / metabolism*
  • MyoD Protein
  • Oligonucleotides / metabolism
  • Polymerase Chain Reaction
  • Protein Conformation
  • Recombinant Fusion Proteins / metabolism
  • Repetitive Sequences, Nucleic Acid
  • TCF Transcription Factors
  • Templates, Genetic
  • Transcription Factor 7-Like 1 Protein
  • Transcription Factors*

Substances

  • DNA-Binding Proteins
  • Macromolecular Substances
  • Muscle Proteins
  • MyoD Protein
  • Oligonucleotides
  • Recombinant Fusion Proteins
  • TCF Transcription Factors
  • Transcription Factor 7-Like 1 Protein
  • Transcription Factors
  • DNA
  • Glutathione Transferase