SpDamID: Marking DNA Bound by Protein Complexes Identifies Notch-Dimer Responsive Enhancers.
SpDamID: Marking DNA Bound by Protein Complexes Identifies Notch-Dimer Responsive Enhancers.
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SPDAMID：通过蛋白质复合物结合的DNA标记可识别Notch-Dimer响应式增强子。
DOI：
10.1016/j.molcel.2015.07.008
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发表时间：
2015-08-20
期刊：
影响因子：
16
通讯作者：
Kopan R
中科院分区：
文献类型：
--
作者：
Hass MR;Liow HH;Chen X;Sharma A;Inoue YU;Inoue T;Reeb A;Martens A;Fulbright M;Raju S;Stevens M;Boyle S;Park JS;Weirauch MT;Brent MR;Kopan R
我们开发了Split DamID（SpDamID），一种DamID的蛋白质互补版本，用于标记体内通过相互作用或并列转录因子结合的基因组DNA。将DAM（DNA腺嘌呤甲基转移酶）的非活性半体融合至待查询的蛋白质对，相互作用或邻近使DAM能够重建和腺嘌呤在GATC中的甲基化。诱导型SpDamID用于分析Notch介导的转录激活。我们证明了Notch复合物标记RBP位点广泛分布在整个基因组中，并表明这些复合物的一个子集招募MAML和p300在响应Notch信号时发生染色质可及性的变化。SpDamID区分单体和二聚体结合，从而允许识别Notch二聚体使用的半位点基序。与SpDamID偶联的Notch增强子的基序富集揭示了Notch和Runx 1对调控序列的共靶向。SpDamID是一种灵敏而强大的工具，可以在全基因组水平上动态分析组合蛋白质-DNA事务。
We developed Split DamID (SpDamID), a protein complementation version of DamID, to mark genomic DNA bound in vivo by interacting or juxtapositioned transcription factors. Inactive halves of DAM (DNA Adenine Methyltransferase) were fused to protein pairs to be queried Interaction or proximity enabled DAM reconstitution and methylation of adenine in GATC. Inducible SpDamID was used to analyze Notch-mediated transcriptional activation. We demonstrate that Notch complexes label RBP sites broadly across the genome, and show that a subset of these complexes that recruit MAML and p300 undergo changes in chromatin accessibility in response to Notch signaling. SpDamID differentiates between monomeric and dimeric binding thereby allowing for identification of half-site motifs used by Notch dimers. Motif enrichment of Notch enhancers coupled with SpDamID reveals co-targeting of regulatory sequences by Notch and Runx1. SpDamID represents a sensitive and powerful tool that enables dynamic analysis of combinatorial protein-DNA transactions at a genome-wide level.