Herman Jean-Paul, Jullien Nicolas, Guillen Séverine, Enjalbert Alain, Pellegrini Isabelle, Franc Jean-Louis
Centre National de la Recherche Scientifique, CRN2M-Unité Mixte de Recherche 7286, F-13344 Marseille, France.
Mol Endocrinol. 2012 Aug;26(8):1455-63. doi: 10.1210/me.2011-1308. Epub 2012 May 25.
The pituitary transcription factor POU1F1 is required for the differentiation of lactotrope, thyrotrope, and somatotrope cells. Its expression is maintained in the adult and is crucial for the expression of prolactin, GH, and TSHβ-subunit. Different studies indicated that POU1F1 could also have other functions in these cells. The identification of new targets of this factor could be useful to obtain a better understanding of these functions. To address this question we combined data obtained from expression microarrays and from chromatin immunoprecipitation (ChIP)-chips. Gene expression microarray assays were used to detect genes that have their expression modified in somatolactotrope GH4C1 cells by the expression of a dominant-negative form of POU1F1, POU1F1(R271W), and led to the identification of 1346 such genes. ChIP-chip experiments were performed from mouse pituitaries and identified 1671 POU1F1-binding sites in gene-promoter regions. Intersecting the gene expression and the ChIP-chip data yielded 121 potential new direct targets. The initial set of 1346 genes identified using the microarrays, as well as the 121 potential new direct targets, were analyzed with DAVID bioinformatics resource for gene ontology term enrichment and cluster. This analysis revealed enrichment in different terms related to protein synthesis and transport, to apoptosis, and to cell division. The present study represents an integrative genome-wide approach to identify new target genes of POU1F1 and downstream networks controlled by this factor.
垂体转录因子POU1F1是促乳素细胞、促甲状腺素细胞和生长激素细胞分化所必需的。其表达在成年期得以维持,对催乳素、生长激素和促甲状腺激素β亚基的表达至关重要。不同研究表明,POU1F1在这些细胞中可能还具有其他功能。鉴定该因子的新靶点可能有助于更好地理解这些功能。为解决这个问题,我们整合了从表达微阵列和染色质免疫沉淀(ChIP)芯片获得的数据。基因表达微阵列分析用于检测在生长激素促乳素细胞GH4C1中,因显性负性形式的POU1F1即POU1F1(R271W)的表达而使其表达发生改变的基因,从而鉴定出1346个此类基因。ChIP芯片实验在小鼠垂体中进行,在基因启动子区域鉴定出1671个POU1F1结合位点。将基因表达数据与ChIP芯片数据进行交叉分析,得到121个潜在的新直接靶点。使用微阵列鉴定出的最初一组1346个基因,以及121个潜在的新直接靶点,利用DAVID生物信息学资源进行基因本体术语富集和聚类分析。该分析揭示了与蛋白质合成和转运、细胞凋亡及细胞分裂相关的不同术语的富集情况。本研究代表了一种全基因组整合方法,用于鉴定POU1F1的新靶基因以及受该因子控制的下游网络。