Miller C P, McGehee R E, Habener J F
Laboratory of Molecular Endocrinology, Massachusetts General Hospital, Boston.
EMBO J. 1994 Mar 1;13(5):1145-56. doi: 10.1002/j.1460-2075.1994.tb06363.x.
We describe the cloning from a rat islet somatostatin-producing cell line of a 1.4 kb cDNA encoding a new homeoprotein, IDX-1 (islet/duodenum homeobox-1), with close sequence similarity to the Drosophila melanogaster homeobox protein Antennapedia (Antp) and the Xenopus laevis endoderm-specific homeoprotein XlHbox8. Analyses of IDX-1 mRNA and protein in rat tissues show that IDX-1 is expressed in pancreatic islets and ducts and in the duodenum. In electrophoretic mobility shift assays IDX-1 binds to three sites in the 5' flanking region of the rat somatostatin gene. In co-transfection experiments IDX-1 transactivates reporter constructs containing somatostatin promoter sequences, and mutation of the IDX-1 binding sites attenuates transactivation. Reverse transcription-polymerase chain reaction of islet RNA using degenerate amplimers for mRNAs encoding homeoproteins indicates that IDX-1 is the most abundant of 12 different Antp-like homeodomain mRNAs expressed in adult rat islets. The pattern of expression, relative abundance and transcriptional regulatory activity suggests that IDX-1 may be involved in the regulation of islet hormone genes and in cellular differentiation in the endocrine pancreas and the duodenum.
我们描述了从大鼠胰岛中产生生长抑素的细胞系克隆出一个1.4 kb的cDNA,该cDNA编码一种新的同源结构域蛋白IDX-1(胰岛/十二指肠同源框-1),它与果蝇的同源框蛋白触角足蛋白(Antp)以及非洲爪蟾内胚层特异性同源结构域蛋白XlHbox8具有高度的序列相似性。对大鼠组织中IDX-1 mRNA和蛋白的分析表明,IDX-1在胰腺胰岛、导管以及十二指肠中表达。在电泳迁移率变动分析中,IDX-1与大鼠生长抑素基因5'侧翼区域的三个位点结合。在共转染实验中,IDX-1可反式激活含有生长抑素启动子序列的报告基因构建体,并且IDX-1结合位点的突变会减弱反式激活作用。使用编码同源结构域蛋白的mRNA的简并引物对胰岛RNA进行逆转录-聚合酶链反应表明,IDX-1是成年大鼠胰岛中表达的12种不同的类Antp同源结构域mRNA中含量最丰富的。其表达模式、相对丰度和转录调控活性表明,IDX-1可能参与胰岛激素基因的调控以及内分泌胰腺和十二指肠中的细胞分化。