Single-molecule regulatory architectures captured by chromatin fiber sequencing
Single-molecule regulatory architectures captured by chromatin fiber sequencing
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DOI：
10.1126/science.aaz1646
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发表时间：
2020-06-26
期刊：
影响因子：
56.9
通讯作者：
Stamatoyannopoulos, John A.
中科院分区：
文献类型：
--
作者：
Stergachis, Andrew B.;Debo, Brian M.;Stamatoyannopoulos, John A.
基因调控主要是在单个线性染色质分子水平上决定的，然而我们目前对顺式调控结构的理解源于对大量不同分子的碎片化采样。我们开发了一种使用非特异性DNA N-6-腺嘌呤甲基转移酶将单个染色质纤维的结构精确地模版到其复合DNA模板上的方法。染色质模板的单分子长读测序能够以核苷酸分辨率读出几千碱基染色质纤维(Fibre-Seq)的初级结构。Fibre-Seq揭示了单个染色质纤维线性组织中广泛存在的可塑性，并阐明了指导调控DNA激活、相邻调控元件的协调激活、单分子核小体定位和单分子转录因子占据的原理。我们的方法和结果为基因调控的主要结构开辟了新的前景。
Gene regulation is chiefly determined at the level of individual linear chromatin molecules, yet our current understanding of cis-regulatory architectures derives from fragmented sampling of large numbers of disparate molecules. We developed an approach for precisely stenciling the structure of individual chromatin fibers onto their composite DNA templates using nonspecific DNA N-6-adenine methyltransferases. Single-molecule long-read sequencing of chromatin stencils enabled nucleotide-resolution readout of the primary architecture of multikilobase chromatin fibers (Fiber-seq). Fiber-seq exposed widespread plasticity in the linear organization of individual chromatin fibers and illuminated principles guiding regulatory DNA actuation, the coordinated actuation of neighboring regulatory elements, single-molecule nucleosome positioning, and single-molecule transcription factor occupancy. Our approach and results open new vistas on the primary architecture of gene regulation.