Genomic Analysis Reveals Distinct Mechanisms and Functional Classes of SOX10-Regulated Genes in Melanocytes [ChIP-Seq]

We performed ChIP-Seq analysis of SOX10, histone H3 lysine 27 acetylation (H3K27ac) and H3K27 trimethylation (H3K27me3) in melanocytes to profile the genomic binding sites of SOX10 and the chromatin landscape. In parallel, we generated Sox10 haploinsufficient cell lines using gene knockout approaches and conducted microarray gene expression analysis to identify functional gene targets of SOX10 transcriptional regulation in melanocytes. We demonstrate that SOX10 predominantly engages “open” chromatin, binds to melanocyte enhancer elements and plays a central role in transcriptional activation and repression of functionally distinct classes of genes. Furthermore, we identified cis-regulatory sequence motifs of putative co-regulatory transcription factors that define SOX10-activated and SOX10-repressed target genes. Our results uncover novel mechanisms and roles of SOX10 in global transcriptional regulation of diverse regulatory pathways in the melanocyte lineage. Overall design: ChIP-seq profiling of SOX10, H3K27ac, and H3K27me3 in the mouse melanocyte cell line melan-Ink4a-Arf-1 (melan-a).

本研究对黑素细胞中的SOX10、组蛋白H3赖氨酸27乙酰化（H3K27ac）及组蛋白H3赖氨酸27三甲基化（H3K27me3）开展染色质免疫共沉淀测序（ChIP-Seq）分析，以解析SOX10的基因组结合位点与染色质景观。与此同时，本研究通过基因敲除技术构建了Sox10单倍剂量不足细胞系，并开展基因芯片表达谱分析，以鉴定黑素细胞中SOX10转录调控的功能性靶基因。研究证实，SOX10主要结合于“开放”染色质，可靶向结合黑素细胞增强子元件，并在功能迥异的基因类别的转录激活与抑制过程中发挥核心作用。此外，本研究还鉴定出可区分SOX10激活型与SOX10抑制型靶基因的推定共调控转录因子顺式调控序列基序。本研究结果揭示了SOX10在黑素细胞谱系多种调控通路的全局转录调控中的全新机制与功能。实验整体设计：对小鼠黑素细胞系melan-Ink4a-Arf-1（简称melan-a）中的SOX10、H3K27ac及H3K27me3开展染色质免疫共沉淀测序分析。