Accurate nanoscale flexibility measurement of DNA and DNA-protein complexes by atomic force microscopy in liquid

Divakaran Murugesapillai, Serge Bouaziz, L. James Maher, Nathan E. Israeloff, Craig E. Cameron, Mark C. Williams

Research output: Contribution to journalArticlepeer-review

18 Scopus citations

Abstract

The elasticity of double-stranded DNA (dsDNA), as described by its persistence length, is critical for many biological processes, including genomic regulation. A persistence length value can be obtained using atomic force microscopy (AFM) imaging. However, most AFM studies have been done by depositing the sample on a surface using adhesive ligands and fitting the contour to a two-dimensional (2D) wormlike chain (WLC) model. This often results in a persistence length measurement that is different from the value determined using bulk and single molecule methods. We describe a method for obtaining accurate three-dimensional (3D) persistence length measurements for DNA and DNA-protein complexes by using a previously developed liquid AFM imaging method and then applying the 3D WLC model. To demonstrate the method, we image in both air and liquid several different dsDNA constructs and DNA-protein complexes that both increase (HIV-1 Vpr) and decrease (yeast HMO1) dsDNA persistence length. Fitting the liquid AFM-imaging contour to the 3D WLC model results in a value in agreement with measurements obtained in optical tweezers experiments. Because AFM also allows characterization of local DNA properties, the ability to correctly measure global flexibility will strongly increase the impact of measurements that use AFM imaging.

Original languageEnglish (US)
Pages (from-to)11327-11337
Number of pages11
JournalNanoscale
Volume9
Issue number31
DOIs
StatePublished - Aug 21 2017

All Science Journal Classification (ASJC) codes

  • Materials Science(all)

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