Measuring shape-dependent looping probability of DNA.

Abstract:

:Recently, several studies have shown that short doubled-stranded DNA (dsDNA) loops more readily than the wormlike chain model predicts. In most of these experiments, the intrinsic bendedness of dsDNA, which in theory can dramatically influence looping dynamics, was either avoided or unaccounted for. To investigate the effect of the shape of dsDNA on looping dynamics, we characterized the shapes of several synthetic dsDNA molecules of equal length but different sequences using gel electrophoresis. We then measured their looping rates using a FRET (Förster resonance energy transfer)-based assay and extracted the looping probability density known as the J factor (jM). We also used, for comparison, several dinucleotide angular parameter sets derived from the observed electrophoretic mobility to compute the jM predicted by the wormlike chain model. Although we found a strong correlation between curvature and jM, the measured jM was higher than most dinucleotide model predictions. This result suggests that it is difficult to reconcile the looping probability with the observed gel mobility within the wormlike chain model and underscores the importance of determining the intrinsic shape of dsDNA for proper theoretical analysis.

journal_name

Biophys J

journal_title

Biophysical journal

authors

Le TT,Kim HD

doi

10.1016/j.bpj.2013.03.029

subject

Has Abstract

pub_date

2013-05-07 00:00:00

pages

2068-76

issue

9

eissn

0006-3495

issn

1542-0086

pii

S0006-3495(13)00368-8

journal_volume

104

pub_type

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