Unit

UNIT 21.4 DNase I and Hydroxyl Radical Characterization of Chromatin Complexes

  1. Joseph M. Vitolo,
  2. Christophe Thiriet,
  3. Jeffrey J. Hayes

Published Online: 1 MAY 2001

DOI: 10.1002/0471142727.mb2104s48

Current Protocols in Molecular Biology

Current Protocols in Molecular Biology

How to Cite

Vitolo, J. M., Thiriet, C. and Hayes, J. J. 2001. DNase I and Hydroxyl Radical Characterization of Chromatin Complexes. Current Protocols in Molecular Biology. 48:21.4:21.4.1–21.4.9.

Author Information

  1. University of Rochester Medical Center, Rochester, New York

Publication History

  1. Published Online: 1 MAY 2001
  2. Published Print: OCT 1999

Abstract

The native chromatin complex within most eukaryotic nuclei is very difficult to study by biochemical means, so researchers have developed methods for studying smaller portions of the complex. This unit details the use of DNase I and hydroxyl radicals to characterize histone-DNA interactions within such portions of the complex. DNase I digestion can be used to determine what regions of a DNA segment are intimately associated with the core histone proteins and what regions are more like naked DNA (i.e., linker DNA within the nucleosomal repeat). The finer deatils of histone-DNA interactions and DNA structure within these complexes is best characterized by digestion with the hydroxyl radical. Both reagents may be used to assess the degree and homogeneity of rotational and translational positioning within isolated chromatin complexes.