Associative behavior of the histone (H3-H4)2 tetramer: dependence on ionic environment.
Mixtures of histones H3 and H4 were examined by analytical ultracentrifugation and circular dichroism to determine their association behavior and secondary structure content in high and low ionic strength solvents containing chloride, phosphate, or sulfate. H3 and H4 were also cross-linked by using DSP in order to directly trap any intermolecular interactions occurring in solution. While H3 and H4 can exist as an H3-H4 dimer under limited conditions, they behave as a stable (H3-H4)2 tetramer under most conditions, particularly those which are physiologically relevant. In chloride-containing solutions, the equilibrium between H3-H4 and (H3-H4)2 is responsive to changes in ionic strength and paralleled by large changes in alpha-helicity. In sulfate- and phosphate-containing solutions, the equilibrium is again governed by ionic strength, but there are no significant changes in secondary structure accompanying shifts in the equilibrium. Small oligomers can be formed in the presence of sulfate and phosphate and trapped by the cross-linking reagent; these oligomers are much smaller than those formed in chloride-containing solutions. However, addition of the H2A-H2B dimer into the system prevents aggregation of the (H3-H4)2 tetramer by acting as a "molecular cap" and thus regulating the assembly pathway toward the formation of tripartite octamers. The observed assembly of H3 and H4 into a stable, tetrameric complex supports the concept of the core histone octamer having a tripartite organization in solution rather than being organized as two heterotypic tetramers.
Baxevanis
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AD
Department of Biology, Johns Hopkins University, Baltimore, Maryland 21218.
Godfrey
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JE
Moudrianakis
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EN
D000818:Animals; D002645:Chickens; D002712:Chlorides; D002942:Circular Dichroism; D003432:Cross-Linking Reagents; D004912:Erythrocytes / Q000737:chemistry; D006657:Histones / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D009994:Osmolar Concentration; D010710:Phosphates; D011487:Protein Conformation; D013329:Structure-Activity Relationship; D013431:Sulfates; D014461:Ultracentrifugation
D002712:Chlorides; D003432:Cross-Linking Reagents; D006657:Histones; D010710:Phosphates; D013431:Sulfates