Structure and function of the mouse insulin-like growth factor binding protein 5 gene promoter.
Structure and function of the mouse insulin-like growth factor binding protein 5 gene promoter.
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小鼠胰岛素样生长因子结合蛋白5基因启动子的结构和功能。
DOI：
10.1089/dna.1995.14.241
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发表时间：
1995
期刊：
影响因子：
--
通讯作者：
Rotwein,P
中科院分区：
文献类型：
--
作者：
Kou,K;Mittanck,DW;Fu,C;Rotwein,P
胰岛素样生长因子I和II(IGF-I和IGF-II)的作用受一个或多个分泌型IGF结合蛋白(IGFBPs)家族的相互作用调节。IGFBP-5是已知的六种IGFBPs中最保守的一种，是一种由252个氨基酸组成的蛋白质，已被证明既能增强IGF的作用，也能抑制IGF的作用。在以前的研究中，我们已经克隆和鉴定了小鼠IGFBP-5基因，并证明了它在成年小鼠不同组织和啮齿动物胚胎发育过程中以分级模式表达。在这份报告中，我们描述了IGFBP-5基因启动子的初步分析。通过瞬时基因转移研究，我们发现小鼠IGFBP-5基因5‘侧翼区31~4,100bp的DNA片段具有定向特异性活性，指导异源报告基因荧光素酶在Hep G2细胞中的表达。只有156个碱基的5‘-侧翼序列的DNA片段介导了60%以上的最大启动子活性，而且含有TATA盒和外显子1的前120个碱基的片段仍具有一定的启动子功能。在高活性的156bp区域内，我们发现了从−70到−34的37bp的片段，该片段在DNaseI足迹和凝胶迁移率改变实验中显示出与Hep G2核蛋白提取物的特异性结合。足迹区域在大鼠和人类的IGFBP-5基因中几乎完全保守，至少占完整启动子活性的70%，这一序列中微小的内部缺失对启动子功能的有害后果就是明证。综上所述，我们的结果表明，小鼠IGFBP-5基因启动子具有简单的结构，并为研究IGFBP-5基因在发育过程中和成人中转录的多因素调控机制奠定了基础。
The actions of insulin-like growth factors I and II (IGF-I and -II) are modulated by interactions with one or more of a family of secreted IGF binding proteins (IGFBPs). IGFBP-5, the most conserved of the six known IGFBPs, is a 252-amino-acid protein that has been shown both to potentiate and inhibit IGF action. In previous studies, we have cloned and characterized the mouse IGFBP-5 gene and demonstrated that it is expressed in a hierarchical pattern in different adult mouse tissues and during rodent embryonic development. In this report, we describe the initial analysis of the IGFBP-5 gene promoter. By transient gene transfer studies, we show the orientation-specific activity of DNA fragments containing from 31 to 4,100 bp from the 5′-flanking region of the mouse IGFBP-5 gene in directing expression of the heterologous reporter gene luciferase in Hep G2 cells. DNA fragments with only 156 bp of 5′-flanking sequence mediated over 60% of maximal promoter activity, and a segment containing the TATA box and the first 120 bp of exon 1 still conferred some promoter function. Within the highly active 156-bp region, we identified a 37-bp segment from −70 to −34 that exhibited specific binding in DNase I footprinting and gel-mobility shift experiments with Hep G2 nuclear protein extracts. The footprinted region, which is almost completely conserved in the rat and human IGFBP-5 genes, was responsible for at least 70% of the activity of the intact promoter, as evidenced by the deleterious consequences of small internal deletions within this sequence on promoter function. Taken together, our results demonstrate that the mouse IGFBP-5 gene promoter has a simple structure, and defines a framework for examining the mechanisms responsible for the multifactorial regulation of IGFBP-5 gene transcription during development and in the adult.
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影响因子：
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作者：
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DOI：
--
发表时间：
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期刊：
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影响因子：
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作者：
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DOI：
--
发表时间：
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期刊：
The Journal of biological chemistry
影响因子：
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作者：
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