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

Journal Article · Tue Apr 21 04:00:00 EDT 1987 · Biochemistry; (United States)

DOI:https://doi.org/10.1021/bi00382a037· OSTI ID:5514782

Jackson, V

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.

• Research Organization: Medical College of Wisconsin, Milwaukee
• OSTI ID: 5514782
• Journal Information: Biochemistry; (United States), Journal Name: Biochemistry; (United States) Vol. 26:8; ISSN BICHA
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
AMINO ACIDS
ANIMAL CELLS
BIOSYNTHESIS
CARBON 13
CARBON ISOTOPES
CARBOXYLIC ACIDS
CHEMICAL REACTIONS
CHROMATIN
CROSS-LINKING
DEUTERIUM COMPOUNDS
ELECTROPHORESIS
EVEN-ODD NUCLEI
HISTONES
HYDROGEN COMPOUNDS
ISOTOPES
LABELLED COMPOUNDS
LIGHT NUCLEI
NITROGEN 15
NITROGEN ISOTOPES
NUCLEI
NUCLEOPROTEINS
NUCLEOSOMES
ODD-EVEN NUCLEI
ORGANIC ACIDS
ORGANIC COMPOUNDS
POLYMERIZATION
PROTEINS
STABLE ISOTOPES
SYNTHESIS
TRITIUM COMPOUNDS