Single molecule visualization and characterization of Sox2-Pax6 complex formation on a regulatory DNA element using a DNA origami frame

Nano Lett. 2014 May 14;14(5):2286-92. doi: 10.1021/nl4044949. Epub 2014 Apr 4.

Abstract

We report the use of atomic force microscopy (AFM) to study Sox2-Pax6 complex formation on the regulatory DNA element at a single molecule level. Using an origami DNA scaffold containing two DNA strands with different levels of tensile force, we confirmed that DNA bending is necessary for Sox2 binding. We also demonstrated that two transcription factors bind cooperatively by observing the increased occupancy of Sox2-Pax6 on the DNA element compared to that of Sox2 alone.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • DNA / chemistry
  • DNA-Binding Proteins / chemistry
  • DNA-Binding Proteins / ultrastructure*
  • Eye Proteins / chemistry
  • Eye Proteins / ultrastructure*
  • Homeodomain Proteins / chemistry
  • Homeodomain Proteins / ultrastructure*
  • Microscopy, Atomic Force
  • Multiprotein Complexes / chemistry
  • Multiprotein Complexes / ultrastructure*
  • Nanotechnology / methods
  • PAX6 Transcription Factor
  • Paired Box Transcription Factors / chemistry
  • Paired Box Transcription Factors / ultrastructure*
  • Protein Binding
  • Repressor Proteins / chemistry
  • Repressor Proteins / ultrastructure*
  • SOXB1 Transcription Factors / chemistry
  • SOXB1 Transcription Factors / ultrastructure*

Substances

  • DNA-Binding Proteins
  • Eye Proteins
  • Homeodomain Proteins
  • Multiprotein Complexes
  • PAX6 Transcription Factor
  • Paired Box Transcription Factors
  • Repressor Proteins
  • SOXB1 Transcription Factors
  • DNA