DNA curtains: novel tools for imaging protein-nucleic acid interactions at the single-molecule level

Methods Cell Biol. 2014:123:217-34. doi: 10.1016/B978-0-12-420138-5.00012-4.

Abstract

Interactions between proteins and nucleic acids are at the molecular foundations of most key biological processes, including DNA replication, genome maintenance, the regulation of gene expression, and chromosome segregation. A complete understanding of these types of biological processes requires tackling questions with a range of different techniques, such as genetics, cell biology, molecular biology, biochemistry, and structural biology. Here, we describe a novel experimental approach called "DNA curtains" that can be used to complement and extend these more traditional techniques by providing real-time information about protein-nucleic acid interactions at the level of single molecules. We describe general features of the DNA curtain technology and its application to the study of protein-nucleic acid interactions in vitro. We also discuss some future developments that will help address crucial challenges to the field of single-molecule biology.

Keywords: Lipid bilayer; Nanofabrication; One-dimensional diffusion; Single molecule; Single-stranded DNA; Total internal reflection fluorescence microscopy; Translocation.

Publication types

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

MeSH terms

  • Biosensing Techniques / instrumentation
  • Biosensing Techniques / methods
  • DNA-Binding Proteins / chemistry*
  • Diffusion
  • Enzyme Assays
  • Immobilized Nucleic Acids / chemistry*
  • Lipid Bilayers / chemistry
  • Microfluidic Analytical Techniques
  • Microscopy, Fluorescence
  • Protein Binding

Substances

  • DNA-Binding Proteins
  • Immobilized Nucleic Acids
  • Lipid Bilayers