Human ISWI chromatin-remodeling complexes sample nucleosomes via transient binding reactions and become immobilized at active sites
Human ISWI chromatin-remodeling complexes sample nucleosomes via transient binding reactions and become immobilized at active sites
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DOI：
10.1073/pnas.1003438107
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发表时间：
2010-11-16
影响因子：
11.1
通讯作者：
Rippe, Karsten
中科院分区：
文献类型：
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作者：
Erdel, Fabian;Schubert, Thomas;Rippe, Karsten
染色质重塑复合体能够以一种依赖于ATP的方式将核小体沿DNA移位。在这里，我们用荧光显微镜/光谱学的方法研究了人ISWI家族成员Snf2h、Snf21、催化失活的Snf21+13剪接变异体以及辅助Acf1亚基在活人和小鼠细胞中的自发荧光蛋白结构。除了在U2OS细胞系中未检测到的Snf21外，在核浓度为0.14~0.83mU时，内源性ISWI蛋白含量丰富。蛋白质相互作用分析表明，多聚体Snf2 h与Acf1结合形成异源四聚体或更高阶的ACF复合体。在G1/2细胞周期中，Snf2、SnF2、Sn
Chromatin remodeling complexes can translocate nucleosomes along the DNA in an ATP-dependent manner. Here, we studied autofluorescent protein constructs of the human ISWI family members Snf2H, Snf2L, the catalytically inactive Snf2L+13 splice variant, and the accessory Acf1 subunit in living human and mouse cells by fluorescence microscopy/spectroscopy. Except for Snf2L, which was not detected in the U2OS cell line, the endogenous ISWI proteins were abundant at nuclear concentrations between 0.14 and 0.83 mu M. A protein interaction analysis showed the association of multimeric Snf2H and Acf1 into a heterotetramer or higher-order ACF complex. During the G1/2 cell cycle phase, Snf2H and Snf2L displayed average residence times