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Isolation of newly replicated chromatin by using shallow metrizamide gradients

Murphy, Robert F. and Wallace, R. Bruce and Bonner, James (1980) Isolation of newly replicated chromatin by using shallow metrizamide gradients. Proceedings of the National Academy of Sciences of the United States of America, 77 (6). pp. 3336-3340. ISSN 0027-8424.

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The properties of chromatin containing newly synthesized DNA and protein were investigated. A fraction of chromatin enriched in newly replicated DNA was isolated by means of its increased density in metrizamide relative to bulk chromatin. The DNA of this fraction appeared to be packaged into nucleosomes but at a reduced nucleosomal spacing. Although pulse-labeled DNA was present in this dense fraction, nucleosomes labeled with short pulses of arginine or acetate were of normal density. The data presented are consistent with the conclusion that newly replicated DNA is associated with preexisting histones in a short-lived, compact structure, whereas newly synthesized histones are deposited at normal spacing some distance from the replication fork.

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Additional Information:© 1980 by the National Academy of Sciences. Contributed by James F. Bonner, March 24, 1980. We thank Thomas D. Sargent for the gift of the 32P-labeled pBR322 markers and for helpful discussion of this manuscript. This work was supported by U.S. Public Health Service Grant GM-13762. The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. §1734 solely to indicate this fact.
Subject Keywords:nucleosome; histone deposition; histone acetylation
Issue or Number:6
Record Number:CaltechAUTHORS:MURpnas80
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Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:7738
Deposited By: Tony Diaz
Deposited On:01 Aug 2007
Last Modified:02 Oct 2019 23:44

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