AN E-BOX ELEMENT IS REQUIRED FOR THE EXPRESSION OF THE AD4BP GENE, A MAMMALIAN HOMOLOG OF FTZ-F1 GENE, WHICH IS ESSENTIAL FOR ADRENAL AND GONADAL DEVELOPMENT
AN E-BOX ELEMENT IS REQUIRED FOR THE EXPRESSION OF THE AD4BP GENE, A MAMMALIAN HOMOLOG OF FTZ-F1 GENE, WHICH IS ESSENTIAL FOR ADRENAL AND GONADAL DEVELOPMENT
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DOI：
10.1074/jbc.270.13.7453
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发表时间：
1995-03-31
影响因子：
4.8
通讯作者：
MOROHASHI, K
中科院分区：
文献类型：
--
作者：
NOMURA, M;BARTSCH, S;MOROHASHI, K
Ad 4 BP又称SF-1，是一种细胞类型特异性的转录因子，调控所有的类固醇生成P-450基因。最近，对编码Ad 4 BP/SF-1的小鼠ftz-f1基因的靶向破坏已经确定了其在肾上腺和性腺发育以及性分化中的重要功能。作为了解其在类固醇生成组织分化和性腺性别分化所必需的基因激活级联中的作用的第一步，我们分离并鉴定了大鼠ad 4 bp基因。ad 4 bp基因的序列分析表明，另一个核因子ELF也转录从同一基因的选择性启动子使用和剪接。通过瞬时转染实验检测，ad 4 bp基因的启动子在类固醇生成细胞如Y-1肾上腺皮质细胞和I-10睾丸Leydig细胞中显示出活性。通过缺失分析和定点突变，我们在5 ′端上游-82 ~-77 bp处鉴定出一个顺式元件，该顺式元件与共有E盒元件一致，是碱性螺旋环螺旋蛋白的结合位点，凝胶迁移率变化分析显示，在从大鼠睾丸制备的核提取物中，与该E盒结合的因子的量在出生后1周达到最大水平然后急剧下降，在成年大鼠中仅检测到痕量。相反，卵巢中的结合因子在出生后达到最高水平，并在出生后保持其水平。这些双态表达的E盒的结合因子与Ad 4 BP的相关性很好，从而表明，Ad 4 BP的表达是通过这个E盒元件的转录调控。
Ad4BP, also known as SF-1, is a cell type-specific transcription factor regulating all the steroidogenic P-450 genes, Recently, the targeted disruption of the mouse ftz-f1 gene encoding Ad4BP/SF-1 has established its essential function in both adrenal and gonadal development and sexual differentiation. As an initial step toward understanding its role in the cascade of gene activations necessary for the differentiation of the steroidogenic tissues and the sex differentiation of the gonads, we isolated and characterized the rat ad4bp gene. A sequence analysis of the ad4bp gene revealed that another nuclear factor ELF was also transcribed from the same gene by alternative promoter usage and splicing. The promoter of the ad4bp gene showed activities in the steroidogenic cells such as Y-1 adrenocortical cells and I-10 testicular Leydig cells when examined by transient transfection assays. Using deletion analysis and site-directed mutagenesis, we identified a cis-element at the position from -82 bp to -77 bp in the 5'-upstream region, The cis-element was identical to the consensus E box element, which is the binding site for the basic-helix-loop-helix proteins, Gel mobility shift analyses revealed the amount of a binding factor to this E box in the nuclear extract prepared from the rat testes attained a maximal level 1 week after birth and then decreased dramatically thereafter, and only trace amounts were detected in adult rats. In contrast, the binding factor in the ovaries attained a maximal level just after birth and kept its level thereafter. These dimorphic expressions of the binding factor to the E box correlated well with those of Ad4BP, and thus suggested that the expression of Ad4BP is transcriptionally regulated through this E box element.