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Proceedings Paper

Rotational diffusion of nucleosomes: role of the N-terminal histone domains in structural transitions
Author(s): David W. Brown; Enoch W. Small
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Paper Abstract

The rotational diffusion of nucleosome core particles were measured using the fluorescence anisotropy decay of ethidium intercalated into the core particle DNA as a function of ionic strength. The `native' form of the core particle (ionic strength from 0.001 to 0.1 M) has a rotational correlation time ((phi) max) of approximately 170 ns. At higher salt concentrations (phi) max rises slowly from approximately 170 ns at 0.1 M NaCl to a value of approximately 230 ns at 0.35 M NaCl, a point just above physiological ionic strength; we call this change the moderate-salt transition. (phi) max remains constant at approximately 230 ns until the onset of the `high-salt dissociation' which occurs above 0.7 M NaCl. This dissociation begins with the release of H2A-H2B dimers; increasing DNA flexibility in this salt range prohibits accurate measurement of the rotational correlation time beyond this point. Light treatment of the ore particles with trypsin to remove the N-terminal histone domains abolishes the plateau in (phi) max between 0.35 and 0.65 M NaCl. Thus, the moderate-salt transition derives from the release of these N-terminal ends from the body of the core particle. The anisotropy decays show no evidence for DNA release from the core particle at salt concentrations below 0.65 M.

Paper Details

Date Published: 1 April 1992
PDF: 4 pages
Proc. SPIE 1640, Time-Resolved Laser Spectroscopy in Biochemistry III, (1 April 1992); doi: 10.1117/12.58276
Show Author Affiliations
David W. Brown, Oregon State Univ. (United States)
Enoch W. Small, Eastern Washington Univ. (United States)

Published in SPIE Proceedings Vol. 1640:
Time-Resolved Laser Spectroscopy in Biochemistry III
Joseph R. Lakowicz, Editor(s)

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