A global analysis of the complex landscape of isoforms and regulatory networks of p63 in human cells and tissues.
A global analysis of the complex landscape of isoforms and regulatory networks of p63 in human cells and tissues.
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DOI：
10.1186/s12864-015-1793-9
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发表时间：
2015-08-07
期刊：
影响因子：
4.4
通讯作者：
Sinha S
中科院分区：
文献类型：
--
作者：
Sethi I;Romano RA;Gluck C;Smalley K;Vojtesek B;Buck MJ;Sinha S
转录因子p63属于p53/p63/p73家族，在正常上皮发育和分化以及病理状态如鳞状细胞癌中起关键的功能作用。人类TP 63基因位于染色体3q 28上，由两个启动子驱动，产生全长反式激活（TA）和N端截短（ΔN）同种型。此外，C末端的选择性剪接产生额外的α、β、γ和可能的其他几种次要变体。梳理出每种p63变体的表达和生物学功能一直是p63领域的焦点和争论的原因。在这里，我们利用了新兴的基于RNA-Seq的基因组数据集来检查p63亚型在常用的人类细胞系和主要组织和器官中的全局表达谱。与早期的研究一致，我们发现Δ Np 63转录本，主要是Δ Np 63 α亚型，在大多数上皮来源的细胞中表达，如皮肤和口腔组织，乳腺和鳞状细胞癌。相比之下，TAp 63在大多数正常细胞类型和组织中不表达;相反，它在伯基特和弥漫性大B细胞淋巴瘤细胞系的亚组中以中等至高水平选择性表达。我们使用新开发的ΔN和TA特异性抗体，通过Western印迹分析验证了p63亚型的这种差异表达模式。此外，使用人类细胞系，组织和器官的无监督聚类，我们表明Δ Np 63和TAp 63驱动的转录网络涉及非常不同的分子参与者，这可能是它们不同生物学功能的基础。在这项研究中，我们报告全面和全球的表达谱的p63亚型和它们的关系，p53/p73和其他潜在的转录辅助调节。我们整理了部分由ENCODE，FANTOM和Human Protein Atlas等财团产生的公开数据，以描绘Δ Np 63和TAp 63的截然不同的转录组学景观。我们的研究不仅有助于消除流行的神话和争议p63在常用的人类细胞系的表达，但也预示着新的亚型和细胞类型特异性活动的p63。本文的在线版本（doi：10.1186/s12864-015-1793-9）包含补充材料，可供授权用户使用。
The transcription factor p63 belongs to the p53/p63/p73 family and plays key functional roles during normal epithelial development and differentiation and in pathological states such as squamous cell carcinomas. The human TP63 gene, located on chromosome 3q28 is driven by two promoters that generate the full-length transactivating (TA) and N-terminal truncated (ΔN) isoforms. Furthermore alternative splicing at the C-terminus gives rise to additional α, β, γ and likely several other minor variants. Teasing out the expression and biological function of each p63 variant has been both the focus of, and a cause for contention in the p63 field. Here we have taken advantage of a burgeoning RNA-Seq based genomic data-sets to examine the global expression profiles of p63 isoforms across commonly utilized human cell-lines and major tissues and organs. Consistent with earlier studies, we find ΔNp63 transcripts, primarily that of the ΔNp63α isoforms, to be expressed in most cells of epithelial origin such as those of skin and oral tissues, mammary glands and squamous cell carcinomas. In contrast, TAp63 is not expressed in the majority of normal cell-types and tissues; rather it is selectively expressed at moderate to high levels in a subset of Burkitt’s and diffuse large B-cell lymphoma cell lines. We verify this differential expression pattern of p63 isoforms by Western blot analysis, using newly developed ΔN and TA specific antibodies. Furthermore using unsupervised clustering of human cell lines, tissues and organs, we show that ΔNp63 and TAp63 driven transcriptional networks involve very distinct sets of molecular players, which may underlie their different biological functions. In this study we report comprehensive and global expression profiles of p63 isoforms and their relationship to p53/p73 and other potential transcriptional co-regulators. We curate publicly available data generated in part by consortiums such as ENCODE, FANTOM and Human Protein Atlas to delineate the vastly different transcriptomic landscapes of ΔNp63 and TAp63. Our studies help not only in dispelling prevailing myths and controversies on p63 expression in commonly used human cell lines but also augur new isoform- and cell type-specific activities of p63. The online version of this article (doi:10.1186/s12864-015-1793-9) contains supplementary material, which is available to authorized users.