Single-Molecule Dynamics of Enhanceosome Assembly in Embryonic Stem Cells

被引：442

作者：

Chen, Jiji ^{[1
]}

Zhang, Zhengjian ^{[3
]}

Li, Li ^{[3
]}

Chen, Bi-Chang ^{[3
]}

Revyakin, Andrey ^{[1
]}

Hajj, Bassam ^{[1
,5
]}

Legant, Wesley ^{[3
]}

Dahan, Maxime ^{[1
,5
]}

Lionnet, Timothee ^{[1
]}

Betzig, Eric ^{[3
]}

Tjian, Robert ^{[1
,3
,4
]}

Liu, Zhe ^{[1
,2
]}

机构：

[1] Howard Hughes Med Inst, Janelia Farm Res Campus, Transcript Imaging Consortium, Ashburn, VA 20147 USA

[2] Howard Hughes Med Inst, Janelia Farm Res Campus, Junior Fellow Program, Ashburn, VA 20147 USA

[3] Howard Hughes Med Inst, Janelia Farm Res Campus, Ashburn, VA 20147 USA

[4] Univ Calif Berkeley, CIRM Ctr Excellence, LKS Biomed & Hlth Sci Ctr, Berkeley, CA 94720 USA

[5] Inst Curie, CNRS, Lab Physo Chim Curie, Unite Mixte Rech 168, F-75794 Paris, France

来源：

CELL | 2014年 / 156卷 / 06期

关键词：

GENE-EXPRESSION; DNA; DIFFUSION; NETWORK; KINETICS; REGIONS; BINDING;

D O I：

10.1016/j.cell.2014.01.062

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Enhancer-binding pluripotency regulators (Sox2 and Oct4) play a seminal role in embryonic stem (ES) cell-specific gene regulation. Here, we combine in vivo and in vitro single-molecule imaging, transcription factor (TF) mutagenesis, and ChIP-exo mapping to determine how TFs dynamically search for and assemble on their cognate DNA target sites. We find that enhanceosome assembly is hierarchically ordered with kinetically favored Sox2 engaging the target DNA first, followed by assisted binding of Oct4. Sox2/Oct4 follow a trial-and-error sampling mechanism involving 84-97 events of 3D diffusion (3.3-3.7 s) interspersed with brief nonspecific collisions (0.75-0.9 s) before acquiring and dwelling at specific target DNA (12.0-14.6 s). Sox2 employs a 3D diffusion-dominated search mode facilitated by 1D sliding along open DNA to efficiently locate targets. Our findings also reveal fundamental aspects of gene and developmental regulation by fine-tuning TF dynamics and influence of the epigenome on target search parameters.

引用

页码：1274 / 1285

页数：12

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