National Institutes of Health/National Human Genome Research Institute
GM073767
米国
National Institutes of Health/National Human Genome Research Institute
R01GM082893
米国
National Institutes of Health/National Human Genome Research Institute
GM108455
米国
National Institutes of Health/National Human Genome Research Institute
1S10OD020054
米国
引用
ジャーナル: Elife / 年: 2019 タイトル: Cryo-EM structures of remodeler-nucleosome intermediates suggest allosteric control through the nucleosome. 著者: Jean Paul Armache / Nathan Gamarra / Stephanie L Johnson / John D Leonard / Shenping Wu / Geeta J Narlikar / Yifan Cheng / 要旨: The SNF2h remodeler slides nucleosomes most efficiently as a dimer, yet how the two protomers avoid a tug-of-war is unclear. Furthermore, SNF2h couples histone octamer deformation to nucleosome ...The SNF2h remodeler slides nucleosomes most efficiently as a dimer, yet how the two protomers avoid a tug-of-war is unclear. Furthermore, SNF2h couples histone octamer deformation to nucleosome sliding, but the underlying structural basis remains unknown. Here we present cryo-EM structures of SNF2h-nucleosome complexes with ADP-BeF that capture two potential reaction intermediates. In one structure, histone residues near the dyad and in the H2A-H2B acidic patch, distal to the active SNF2h protomer, appear disordered. The disordered acidic patch is expected to inhibit the second SNF2h protomer, while disorder near the dyad is expected to promote DNA translocation. The other structure doesn't show octamer deformation, but surprisingly shows a 2 bp translocation. FRET studies indicate that ADP-BeF predisposes SNF2h-nucleosome complexes for an elemental translocation step. We propose a model for allosteric control through the nucleosome, where one SNF2h protomer promotes asymmetric octamer deformation to inhibit the second protomer, while stimulating directional DNA translocation.
全体 : Cryo-EM structure of singly-bound SNF2h-nucleosome complex at 6.9...
全体
名称: Cryo-EM structure of singly-bound SNF2h-nucleosome complex at 6.9A with SNF2h bound at SHL+2
要素
複合体: Cryo-EM structure of singly-bound SNF2h-nucleosome complex at 6.9A with SNF2h bound at SHL+2
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超分子 #1: Cryo-EM structure of singly-bound SNF2h-nucleosome complex at 6.9...
超分子
名称: Cryo-EM structure of singly-bound SNF2h-nucleosome complex at 6.9A with SNF2h bound at SHL+2 タイプ: complex / ID: 1 / 親要素: 0
由来(天然)
生物種: Xenopus laevis (アフリカツメガエル)
組換発現
生物種: Escherichia coli (大腸菌) / 組換株: BL21(DE3)
分子量
理論値: 340 kDa/nm
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実験情報
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構造解析
手法
クライオ電子顕微鏡法
解析
単粒子再構成法
試料の集合状態
particle
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試料調製
緩衝液
pH: 7.5
グリッド
モデル: Quantifoil R1.2/1.3 / 材質: COPPER / メッシュ: 400
凍結
凍結剤: ETHANE / チャンバー内湿度: 100 % / チャンバー内温度: 295.15 K / 装置: FEI VITROBOT MARK I 詳細: 2.5 ul of nucleosome-443 SNF2h complexes were applied to a glow discharged Quantifoil holey carbon grid (1.2 um hole size, 400 mesh), blotted in a Vitrobot Mark I (FEI Company) using 6 ...詳細: 2.5 ul of nucleosome-443 SNF2h complexes were applied to a glow discharged Quantifoil holey carbon grid (1.2 um hole size, 400 mesh), blotted in a Vitrobot Mark I (FEI Company) using 6 seconds blotting at 100% humidity, and then plunge-frozen in liquid ethane cooled by liquid nitrogen..