Differences between MyoD DNA binding and activation site requirements revealed by functional random sequence selection

Mol Cell Biol. 1996 Jul;16(7):3893-900. doi: 10.1128/MCB.16.7.3893.


A method has been developed for selecting functional enhancer/promoter sites from random DNA sequences in higher eukaryotic cells. Of sequences that were thus selected for transcriptional activation by the muscle-specific basic helix-loop-helix protein MyoD, only a subset are similar to the preferred in vitro binding consensus, and in the same promoter context an optimal in vitro binding site was inactive. Other sequences with full transcriptional activity instead exhibit sequence preferences that, remarkably, are generally either identical or very similar to those found in naturally occurring muscle-specific promoters. This first systematic examination of the relation between DNA binding and transcriptional activation by basic helix-loop-helix proteins indicates that binding per se is necessary but not sufficient for transcriptional activation by MyoD and implies a requirement for other DNA sequence-dependent interactions or conformations at its binding site.

Publication types

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

MeSH terms

  • 3T3 Cells
  • Animals
  • Base Sequence
  • Binding Sites
  • DNA / chemistry
  • DNA / metabolism*
  • Enhancer Elements, Genetic
  • Flow Cytometry
  • Helix-Loop-Helix Motifs
  • Mice
  • Molecular Sequence Data
  • Mutagenesis, Insertional
  • MyoD Protein / biosynthesis
  • MyoD Protein / chemistry
  • MyoD Protein / metabolism*
  • Promoter Regions, Genetic
  • Recombinant Fusion Proteins / biosynthesis
  • Substrate Specificity
  • Transfection
  • beta-Galactosidase / biosynthesis


  • MyoD Protein
  • Recombinant Fusion Proteins
  • DNA
  • beta-Galactosidase