Single chromatin fiber stretching reveals physically distinct populations of disassembly events.
Eukaryotic DNA is packaged into the cell nucleus as a nucleoprotein complex, chromatin. Despite this condensed state, access to the DNA sequence must occur during gene expression and other essential genetic events. Here we employ optical tweezers stretching of reconstituted chromatin fibers to investigate the release of DNA from its protein-bound structure. Analysis of fiber length increase per unbinding event revealed discrete values of approximately 30 and approximately 60 nm. Furthermore, a loading rate analysis of the disruption forces revealed three individual energy barriers. The heights of these barriers were found to be approximately 20 k(B)T, approximately 25 k(B)T, and approximately 28 k(B)T. For subsequent stretches of the fiber it was found that events corresponding to the approximately 28 k(B)T energy barrier were significantly reduced. No correlation between energy barrier crossed and DNA length release was found. These studies clearly demonstrate that optical tweezers stretching of chromatin provides insight into the energetic penalties imposed by chromatin structure. Furthermore these studies reveal possible pathways via which chromatin may be disrupted during genetic code access.
Pope
L H
LH
Biophysical Techniques, Department of Science and Technology and MESA Institute for Nanotechnology, University of Twente, 7500 AE Enschede, The Netherlands.
Bennink
M L
ML
van Leijenhorst-Groener
K A
KA
Nikova
D
D
Greve
J
J
Marko
J F
JF
D000818:Animals; D001703:Biophysics / Q000295:instrumentation* / Q000379:methods*; D019921:Biotinylation; D002843:Chromatin / Q000737:chemistry*; D004247:DNA / Q000737:chemistry*; D006657:Histones / Q000737:chemistry; D046911:Macromolecular Substances / Q000737:chemistry; D015233:Models, Statistical; D009707:Nucleosomes / Q000737:chemistry; D009865:Oocytes / Q000378:metabolism; D011312:Pressure; D011487:Protein Conformation; D019809:Streptavidin / Q000737:chemistry; D013696:Temperature; D013816:Thermodynamics; D014982:Xenopus laevis
D002843:Chromatin; D006657:Histones; D046911:Macromolecular Substances; D009707:Nucleosomes; D004247:DNA; D019809:Streptavidin