The identification of Hoxc8 target genes
The identification of Hoxc8 target genes
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DOI：
10.1073/pnas.0409700102
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发表时间：
2005-02-15
影响因子：
11.1
通讯作者：
Ruddle, FH
中科院分区：
文献类型：
--
作者：
Lei, HY;Wang, HL;Ruddle, FH
Hox基因编码的转录因子在胚胎发生过程中控制空间格局。迄今为止，Hox基因的下游靶点已被证明难以识别。在这里，我们描述的研究，旨在确定目标基因的控制下的鼠转录因子Hoxc 8。我们使用小鼠16，463基因寡核苷酸微阵列来鉴定其表达被细胞培养（体外）中C57 BL/6 J小鼠胚胎成纤维细胞（MEF）中Hoxc 8过表达改变的mRNA。我们共鉴定了34个基因，其表达变化为2倍或更大：16个基因上调，18个基因下调。大多数基因编码的蛋白质参与关键的生物过程，如细胞粘附，迁移，代谢，凋亡和肿瘤发生。两个基因显示出高水平的调节：（i）分泌型磷蛋白1（Sppl），也称为骨桥蛋白（OPN），下调4.8倍，和（h）卷曲同源物2（果蝇）（Fzd 2）上调4.4倍。染色质免疫沉淀（ChIP）分析证实了OPN启动子和Hoxc 8蛋白1之间的直接相互作用。支持OPN是Hoxc 8的直接转录靶点的观点。
Hox genes encode transcription factors that control spatial patterning during embryogenesis. To date, downstream targets of Hox genes have proven difficult to identify. Here, we describe studies designed to identify target genes under the control of the murine transcription factor Hoxc8. We used a mouse 16,463 gene oligonucleotide microarray to identify mRNAs whose expression was altered by the overexpression of Hoxc8 in C57BL/6J mouse embryo fibroblasts (MEF) in cell culture (in vitro). We identified a total of 34 genes whose expression was changed by 2-fold or greater: 16 genes were up-regulated, and 18 genes were down-regulated. The majority of genes encoded proteins involved in critical biological processes, such as cell adhesion, migration, metabolism, apoptosis, and tumorigenesis. Two genes showed high levels of regulation: (i) secreted phosphoprotein 1 (Sppl), also known as osteopontin (OPN), was down-regulated 4.8-fold, and (h) frizzled homolog 2 (Drosophila) (Fzd2) was up-regulated 4.4-fold. Chromatin immunoprecipitation (ChIP) analysis confirmed the direct interaction between the OPN promoter and Hoxc8 protein 1. n vivo, supporting the view that OPN is a direct transcriptional target of Hoxc8.