核酸特性 结构和功能.ppt

免疫学信息网 19 免疫学信息网 Are Nucleosomes “Positioned?” 1. Sequence at Unique Position 2. Micrococcal Nuclease Digest 3. Restriction Digest 4. Southern Blot Discrete Band Positioning Smear Random Orientation 17 免疫学信息网 Implications of Translational Positioning 18 免疫学信息网 N-ARTKQTARKSTGGKAPRKQLATKAARKSAPATGGVKKPHRYRPGTVALREIRRYQKSTE- H3 N-SGRGKGGKGLGKGGAKRHRKVLRDNIQ - H4 N-SGRGKQGGKTRAKSKTRSSRAGL- H2A N-SDPAKSAPAAKKGSKKAVTKTQKKDGKKRRKSRKES- H2B H3 -ERA-C H4 -RTLYGFGG-C H2A -ELNKLLGGVTIAQGGVLPNIQSVLLPKKTESSKSTKSK-C H2B -PGELAKHAVSEGTKAVTKYTSAK-C Core Core Histones Have Variable Length Tails 20 免疫学信息网 H4 H2B H2A H3 H3 N-terminus H3 N-terminus 21 免疫学信息网 H4 H2B H2A H3 H2B N-terminus 22 免疫学信息网 H4 H2B H2A + + + _ _ _ 23 免疫学信息网 N-SGRGKGGKGLGKGGAKRHRKVLRDNIQ -H4 P Ac Ac Ac Ac N-ARTKQTARKSTGGKAPRKQLATKAARKSAPATGGVKKPHRYRPGTVALREIRRYQKSTE- H3 Core P P 10 28 Ac Ac Ac Ac Ac Ac N-SGRGKQGGKTRAKSKTRSSRAGL-H2A P Ac Ac N-SDPAKSAPAAKKGSKKAVTKTQKKDGKKRRKSRKES-H2B P Ac Ac Ac Ac Ac Ac 23A Newly-synthesized histones M M M M M K79 Active region Inactive Region (heterochromatin, X-inactivation) Active region Mitosis (Aurora B) SSN6/TUP1 免疫学信息网 ??? “Beads on a String” 30 nm Filament 24 Low salt High salt How does chromatin become compacted?? 免疫学信息网 The “Linker” Histone H1/H5 Largest histone (21 kD), less conserved Binds 165 bp: “Chromatosome” Facilitates chromatin compaction 免疫学信息网 Histone H5 reduces salt requirement for chromatin compaction 25 免疫学信息网 Helix III - Major groove binding Loop (Secondary DNA binding site) Globular domain of Histone H5 “winged helix-turn-helix domain” 26 免疫学信息网 N C C C C C C 26 Zhou et al., 1998 Nature 395, 402 免疫学信息网 The Solenoid Model for the 30 nm Fiber 26A Histone H1 免疫学信息网 The 30 nm Chromatin Fiber 5 Compaction ratio = 50 免疫学信息网 Condensins Condensins Are Required for Mitotic Chromosome Condensation Hirano and Mitchison, Cell 79, 449 Condensin- Depleted Mock- Depleted Hirano et al., Cell 89, 511 27 免疫学信息网 Interference Fo