title: Reversible oligonucleosome self-association: dependence on divalent cations and core histone tail domains.
abstract: Regularly spaced nucleosomal arrays equilibrate between unfolded and highly folded conformations in <2 mM MgCl2, and self-associate above 2 mM MgCl2 [Schwarz, P. M., & Hansen, J. C. (1994) J. Biol. Chem. 269, 16284-16289]. Here we use analytical and differential sedimentation techniques to characterize the molecular mechanism and determinants of oligonucleosome self-association. Divalent cations induce self-association of intact nucleosomal arrays by binding to oligonucleosomal DNA and neutralizing its negative charge. Neither linker histones nor H2A/H2B dimers are required for Mg2+ - dependent self-association. However, divalent cations are unable to induce self-association of trypsinized nucleosomal arrays lacking their N- and C-terminal core histone tail domains. This suggests that the H3/H4 tail domains directly mediate oligonucleosome self-association through a non-Coulombic-based mechanism. Self-association occurs independently of whether the oligonucleosome monomers are folded or unfolded. The first step in the self-association pathway is strongly cooperative and produces a soluble association intermediate that sediments approximately 10 times faster than the oligonucleosome monomers. The size of the oligonucleosome polymers increases rapidly as a consequence of small increases in the divalent cation concentration, eventually producing polymeric species that sediment at >> 10 000 S. Importantly, all steps in the self-association pathway are freely reversible upon removal of the divalent cations. Taken together, these data indicate that short oligonucleosome fragments composed of only core histone octamers and DNA possess all of the structural features required to achieve chromosome-level DNA compaction. These findings provide a molecular basis for explaining many of the recently uncovered structural features of interphase and metaphase chromosomal fibers.
authors.affiliation: Department of Biochemistry, The University of Texas Health Science Center at San Antonio, 78284-7760, USA.
authors.forename: P M
authors.initials: PM
authors.lastname: Schwarz
authors.forename: A
authors.initials: A
authors.lastname: Felthauser
authors.forename: T M
authors.initials: TM
authors.lastname: Fletcher
authors.forename: J C
authors.initials: JC
authors.lastname: Hansen
mesh_terms: D000818:Animals; D000838:Anions / Q000494:pharmacology; D001464:Barium / Q000494:pharmacology; D002104:Cadmium / Q000494:pharmacology; D002413:Cations, Divalent / Q000494:pharmacology*; D002414:Cations, Monovalent / Q000494:pharmacology; D002645:Chickens; D003035:Cobalt / Q000494:pharmacology; D004912:Erythrocytes; D006657:Histones / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism*; D007700:Kinetics; D046911:Macromolecular Substances; D008274:Magnesium / Q000494:pharmacology; D015636:Magnesium Chloride / Q000494:pharmacology; D008345:Manganese / Q000494:pharmacology; D009707:Nucleosomes / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure*; D015032:Zinc / Q000494:pharmacology
chemical_list: D000838:Anions; D002413:Cations, Divalent; D002414:Cations, Monovalent; D006657:Histones; D046911:Macromolecular Substances; D009707:Nucleosomes; D002104:Cadmium; D015636:Magnesium Chloride; D001464:Barium; D003035:Cobalt; D008345:Manganese; D008274:Magnesium; D015032:Zinc