Linker histone H1 per se can induce three-dimensional folding of chromatin fiber.
Higher-order architectures of chromosomes play important roles in the regulation of genome functions. To understand the molecular mechanism of genome packing, an in vitro chromatin reconstitution method and a single-molecule imaging technique (atomic force microscopy) were combined. In 50 mM NaCl, well-stretched beads-on-a-string chromatin fiber was observed. However, in 100 mM NaCl, salt-induced interaction between nucleosomes caused partial aggregation. Addition of histone H1 promoted a further folding of the fiber into thicker fibers 20-30 nm in width. Micrococcal nuclease digestion of these thicker fibers produced an approximately 170 bp fragment of nucleosomal DNA, which was approximately 20 bp longer than in the absence of histone H1 ( approximately 150 bp), indicating that H1 is correctly placed at the linker region. The width of the fiber depended on the ionic strength. Widths of 20 nm in 50 mM NaCl became 30 nm as the ionic strength was changed to 100 mM. On the basis of these results, a flexible model of chromatin fiber formation was proposed, where the mode of the fiber compaction changes depending both on salt environment and linker histone H1. The biological significance of this property of the chromatin architecture will be apparent in the closed segments ( approximately 100 kb) between SAR/MAR regions.
Hizume
Kohji
K
Laboratory of Plasma Membrane and Nuclear Signaling, Graduate School of Biostudies, Kyoto University, Kitashirakawa-oiwake-cho, Sakyo-ku, Kyoto 606-8502, Japan.
Yoshimura
Shige H
SH
Takeyasu
Kunio
K
D002843:Chromatin / Q000737:chemistry* / Q000648:ultrastructure; D006367:HeLa Cells; D006657:Histones / Q000302:isolation & purification / Q000502:physiology*; D006801:Humans; D018625:Microscopy, Atomic Force; D009707:Nucleosomes / Q000648:ultrastructure; D009994:Osmolar Concentration; D017510:Protein Folding; D012965:Sodium Chloride / Q000494:pharmacology
D002843:Chromatin; D006657:Histones; D009707:Nucleosomes; D012965:Sodium Chloride