high-throughput sequencing identifies stat3 as the dna-associated factor for p53-nf-κb-complex-dependent gene expression in human heart failure高通量测序识别stat3作为p53-nf-κb-complex-dependent dna-associated因子基因表达在人类心脏衰竭.pdfVIP

Choy et al. Genome Medicine 2010, 2:37 /content/2/6/37 R E S E A R C H Open Access Research High-throughput sequencing identifies STAT3 as the DNA-associated factor for p53-NF-κB-complex-dependent gene expression in human heart failure 1 1 1 1 2 3 3 Mun-Kit Choy , Mehregan Movassagh , Lee Siggens , Ana Vujic , Martin Goddard , Ana Sánchez , Neil Perkins , Nichola Figg1 1 4 1 , Martin Bennett , Jason Carroll and Roger Foo* Abstract Background: 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. Methods: 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. Results: 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