Automated electron microscope tomography of frozen-hydrated chromatin: the irregular three-dimensional zigzag architecture persists in compact, isolated fibers

Journal of Structural Biology
R A HorowitzChristopher L Woodcock

Abstract

The potential of electron microscope tomography as a tool for obtaining three-dimensional (3D) information about large macromolecular assemblies is greatly extended by automation of data collection. With the implementation of automated control of tilting, focusing, and digital image recording described here, tilt series of frozen-hydrated specimens can be collected with the requisite low dose. Long chromatin fibers were prepared in 90 mM monovalent ions to maintain a fully compact conformation, and after vitrification were completely contained within the ice layer. Tilt series of this material were recorded at 5 degrees tilt increments between +60 degrees and -60 degrees, with a cumulative dose of approximately 35 e-/A2 for the series. This extremely low dose data was successfully aligned, then reconstructed by weighted backprojection. The underlying architecture of the fibers is an irregular 3D zigzag of interconnected nucleosomes, with the linker DNA between successive nucleosomes in a largely extended conformation. The visualization of this structural motif within long, frozen-hydrated chromatin fibers at relatively high salt extends our previous studies on small fragments at low ionic strength and is in agreement with the o...Continue Reading

References

Jan 1, 1992·The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society·R A Horowitz, C L Woodcock
Oct 1, 1992·Ultramicroscopy·A J KosterD A Agard
Oct 11, 1984·Nature·T J RichmondA Klug
Oct 1, 1993·Proceedings of the National Academy of Sciences of the United States of America·C L WoodcockN Whitaker
Jan 1, 1993·Journal of Structural Biology·R H Wade, D Chrétien
Jan 1, 1996·Journal of Structural Biology·J P Schroeter, J P Bretaudiere
Jan 1, 1996·Journal of Structural Biology·J C FungD A Agard
Jan 1, 1997·Biophysical Journal·R GrimmW Baumeister
Jul 1, 1995·Trends in Cell Biology·C L Woodcock, R A Horowitz

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Citations

Dec 18, 2001·Journal of Neuroscience Research·G A PerkinsM H Ellisman
Dec 20, 2005·Chromosoma·Rachel A Horowitz-Scherer, Christopher L Woodcock
Feb 18, 2011·Biochemistry and Cell Biology = Biochimie Et Biologie Cellulaire·Heather J Szerlong, Jeffrey C Hansen
Dec 22, 1999·Archives of Histology and Cytology·P GobbiG Mazzotti
May 28, 2005·Proceedings of the National Academy of Sciences of the United States of America·Jian SunTamar Schlick
Nov 25, 1998·Proceedings of the National Academy of Sciences of the United States of America·J BednarC L Woodcock
Apr 29, 2015·FEBS Letters·Guohong Li, Ping Zhu
Nov 28, 2001·Journal of Structural Biology·A S Frangakis, R Hegerl
Jan 4, 2005·Biochemistry. Biokhimii︠a︡·S MarcoJ-L Rigaud
Jun 6, 2003·The Journal of Biological Chemistry·Philippe T GeorgelJeffrey C Hansen

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