High-throughput sequencing identifies STAT3 as the DNA-associated factor for p53-NF-kappaB-complex-dependent gene expression in human heart failure.
High-throughput sequencing identifies STAT3 as the DNA-associated factor for p53-NF-kappaB-complex-dependent gene expression in human heart failure.
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DOI：
10.1186/gm158
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发表时间：
2010-06-14
期刊：
影响因子：
12.3
通讯作者：
Foo R
中科院分区：
文献类型：
--
作者：
Choy MK;Movassagh M;Siggens L;Vujic A;Goddard M;Sánchez A;Perkins N;Figg N;Bennett M;Carroll J;Foo R
DNA调控元件的全基因组图谱及其与转录因子的相互作用可以形成理解人类疾病中的调控回路和基因表达控制的框架，但是这些网络，包括转录因子和DNA结合蛋白，如何形成复合物，与DNA相互作用并调节基因表达仍然是未知的。使用microRNA-21（mir-21），这是在心力衰竭中调节的基因的一个例子，我们进行了染色质免疫沉淀（ChIP）测定，以确定转录因子在该遗传位点的占有率。组织ChIP进一步使用人类心脏和这些转录因子的全基因组的占有率进行了分析，通过高通量测序。我们发现转录因子p53与NF-κB/RELA结合，并利用κ B基序的顺式调控区来控制mir-21的表达。p53在该复合物中表现为辅因子，因为尽管其DNA结合结构域发生突变，突变型p53仍然能够结合RELA和顺式元件，并诱导mir-21表达。在扩张的人心脏中，mir-21上调先前已被证实，p53-RELA复合物也与此顺式元件相关。使用高通量测序，我们分析了患病和对照人类心脏中p53-RELA复合物的全基因组结合位点，发现STAT 3基序的显著过度表达。我们进一步确定STAT 3是p53-RELA复合物与该顺式元件结合以及mir-21表达所必需的。我们的研究结果揭示了转录因子在顺式调控元件处以多分子复合物形式合作控制基因表达的机制。
Genome-wide maps of DNA regulatory elements and their interaction with transcription factors may form a framework for understanding regulatory circuits and gene expression control in human disease, but how these networks, comprising transcription factors and DNA-binding proteins, form complexes, interact with DNA and modulate gene expression remains largely unknown. Using microRNA-21 (mir-21), which is an example of genes that are regulated in heart failure, we performed chromatin immunoprecipitation (ChIP) assays to determine the occupancy of transcription factors at this genetic locus. Tissue ChIP was further performed using human hearts and genome-wide occupancies of these transcription factors were analyzed by high-throughput sequencing. We show that the transcription factor p53 piggy-backs onto NF-κB/RELA and utilizes the κB-motif at a cis-regulatory region to control mir-21 expression. p53 behaves as a co-factor in this complex because despite a mutation in its DNA binding domain, mutant p53 was still capable of binding RELA and the cis-element, and inducing mir-21 expression. In dilated human hearts where mir-21 upregulation was previously demonstrated, the p53-RELA complex was also associated with this cis-element. Using high-throughput sequencing, we analyzed genome-wide binding sites for the p53-RELA complex in diseased and control human hearts and found a significant overrepresentation of the STAT3 motif. We further determined that STAT3 was necessary for the p53-RELA complex to associate with this cis-element and for mir-21 expression. Our results uncover a mechanism by which transcription factors cooperate in a multi-molecular complex at a cis-regulatory element to control gene expression.
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影响因子：
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作者：
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