Detection of Nucleosomal Substructures using Solid-State Nanopores
Detection of Nucleosomal Substructures using Solid-State Nanopores
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DOI：
10.1021/nl301163m
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发表时间：
2012-06-01
期刊：
影响因子：
10.8
通讯作者：
Dekker, Cees
中科院分区：
文献类型：
--
作者：
Soni, Gautam V.;Dekker, Cees
组蛋白聚集在DNA上形成核小体，控制真核细胞染色质的结构和功能。更具体地说，核小体的结构完整性调节基因的表达速率，并作为细胞凋亡的重要早期标志。然而，核小体(亚)结构很难检测和表征。在这里，我们展示了固态纳米孔非常适合于快速且无标记地检测核小体及其组蛋白亚复合体。(核)蛋白质复合体在外加电场的作用下单独穿过纳米孔，从而产生特征的电导阻断，提供有关转位复合体分子大小的定量信息。我们观察到电导阻断和易位时间与核小体亚结构的分子量有系统的依赖关系。这使得能够在单一复合体水平上区分和表征这些蛋白质和DNA-蛋白质复合体。最后，我们展示了区分核小体和二核小体的能力，作为使用纳米孔平台研究染色质阵列的第一步。
Histone proteins assemble onto DNA into nucleosomes that control the structure and function of eukaryotic chromatin. More specifically, the structural integrity of nucleosomes regulates gene expression rates and serves as an important early marker for cell apoptosis. Nucleosomal (sub)structures are however hard to detect and characterize. Here, we show that solid-state nanopores are well suited for fast and label-free detection of nucleosomes and its histone subcomplexes. (Nucleo-)protein complexes are individually driven through the nanopore by an applied electric field, which results in characteristic conductance blockades that provide quantitative information on the molecular size of the translocating complex. We observe a systematic dependence of the conductance blockade and translocation time on the molecular weight of the nucleosomal substructures. This allows discriminating and characterizing these protein and DNA-protein complexes at the single-complex level. Finally, we demonstrate the ability to distinguish nucleosomes and dinucleosomes as a first step toward using the nanopore platform to study chromatin arrays.