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Title: Deposition of newly synthesized histones: new histones H2A and H2B do not deposit in the same nucleosome with new Histones H3 and H4

Abstract

The authors have developed procedures to study histone-histone interactions during the deposition of histones in replicating cells. Cells are labeled for 60 min with dense amino acids, and subsequently, the histones within the nucleosomes are cross-linked into an octameric complex with formaldehyde. These complexes are sedimented to equilibrium in density gradients and octamer and dioctamer complexes separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. With reversal of the cross-link, the distribution of the individual density-labeled histones in the octamer is determined. Newly synthesized H3 and H4 deposits as a tetramer and are associated with old H2A and H2B. Newly synthesized H2A and H2B deposit as a dimer associated with old H2A, H2B, H3, and H4. The significance of these results with respect to the dynamics of histone interactions in the nucleus is discussed. Control experiments are presented to test for artifactual formation of these complexes during preparative procedures. In addition, reconstitution experiments were performed to demonstrate that the composition of these octameric complexes can be determined from their distribution of density gradients.

Authors:
Publication Date:
Research Org.:
Medical College of Wisconsin, Milwaukee
OSTI Identifier:
5514782
Resource Type:
Journal Article
Journal Name:
Biochemistry; (United States)
Additional Journal Information:
Journal Volume: 26:8
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; HISTONES; BIOSYNTHESIS; CROSS-LINKING; LABELLED COMPOUNDS; AMINO ACIDS; ANIMAL CELLS; CARBON 13; DEUTERIUM COMPOUNDS; ELECTROPHORESIS; NITROGEN 15; NUCLEOSOMES; TRITIUM COMPOUNDS; CARBON ISOTOPES; CARBOXYLIC ACIDS; CHEMICAL REACTIONS; CHROMATIN; EVEN-ODD NUCLEI; HYDROGEN COMPOUNDS; ISOTOPES; LIGHT NUCLEI; NITROGEN ISOTOPES; NUCLEI; NUCLEOPROTEINS; ODD-EVEN NUCLEI; ORGANIC ACIDS; ORGANIC COMPOUNDS; POLYMERIZATION; PROTEINS; STABLE ISOTOPES; SYNTHESIS; 550201* - Biochemistry- Tracer Techniques

Citation Formats

Jackson, V. Deposition of newly synthesized histones: new histones H2A and H2B do not deposit in the same nucleosome with new Histones H3 and H4. United States: N. p., 1987. Web. doi:10.1021/bi00382a037.
Jackson, V. Deposition of newly synthesized histones: new histones H2A and H2B do not deposit in the same nucleosome with new Histones H3 and H4. United States. https://doi.org/10.1021/bi00382a037
Jackson, V. 1987. "Deposition of newly synthesized histones: new histones H2A and H2B do not deposit in the same nucleosome with new Histones H3 and H4". United States. https://doi.org/10.1021/bi00382a037.
@article{osti_5514782,
title = {Deposition of newly synthesized histones: new histones H2A and H2B do not deposit in the same nucleosome with new Histones H3 and H4},
author = {Jackson, V},
abstractNote = {The authors have developed procedures to study histone-histone interactions during the deposition of histones in replicating cells. Cells are labeled for 60 min with dense amino acids, and subsequently, the histones within the nucleosomes are cross-linked into an octameric complex with formaldehyde. These complexes are sedimented to equilibrium in density gradients and octamer and dioctamer complexes separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. With reversal of the cross-link, the distribution of the individual density-labeled histones in the octamer is determined. Newly synthesized H3 and H4 deposits as a tetramer and are associated with old H2A and H2B. Newly synthesized H2A and H2B deposit as a dimer associated with old H2A, H2B, H3, and H4. The significance of these results with respect to the dynamics of histone interactions in the nucleus is discussed. Control experiments are presented to test for artifactual formation of these complexes during preparative procedures. In addition, reconstitution experiments were performed to demonstrate that the composition of these octameric complexes can be determined from their distribution of density gradients.},
doi = {10.1021/bi00382a037},
url = {https://www.osti.gov/biblio/5514782}, journal = {Biochemistry; (United States)},
number = ,
volume = 26:8,
place = {United States},
year = {Tue Apr 21 00:00:00 EDT 1987},
month = {Tue Apr 21 00:00:00 EDT 1987}
}