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首页> 外文期刊>Molecular and Cellular Biology >Dioxin receptor and C/EBP regulate the function of the glutathione S-transferase Ya gene xenobiotic response element.
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Dioxin receptor and C/EBP regulate the function of the glutathione S-transferase Ya gene xenobiotic response element.

机译：二恶英受体和C / EBP调节谷胱甘肽S-转移酶Ya基因异源应答元件的功能。

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The rat glutathione S-transferase Ya gene xenobiotic response element (XRE) has both constitutive and xenobiotic-inducible activity. We present evidence that the XRE is regulated by both the constitutive C/EBP transcription factor and the xenobiotic-activated dioxin receptor. A ligand-activated XRE-binding protein was shown to be dioxin receptor by specific antibody immunodepletion and binding of highly purified receptor. Identification of C/EBP alpha as the constitutive binding protein was demonstrated by competition with a C/EBP binding site, protein-DNA cross-linking to determine the molecular weight of the constitutive protein(s), specific antibody immunodepletion, and binding of purified bacterially expressed C/EBP alpha. Mutational analysis of the XRE revealed that the constitutive factor (C/EBP alpha) shares a nearly identical overlapping binding site with the dioxin receptor. In functional testing of the putative C/EBP-XRE interaction, cotransfected C/EBP alpha activated an XRE test promoter in the non-xenobiotic-responsive HeLa cell line. Unexpectedly, cotransfected C/EBP alpha had no effect on basal activity but significantly increased the xenobiotic response of the XRE test promoter in the xenobiotic-responsive, C/EBP-positive HepG2 cell line. Furthermore, inhibition of C/EBP-binding protein(s) in HepG2 cells by transfection of C/EBP oligonucleotides suppressed the xenobiotic response. These results suggest that C/EBP alpha and dioxin receptor recognize the same DNA sequence element and that transcriptional regulation can occur by cooperative interactions between these two transcription factors.
机译：大鼠谷胱甘肽S-转移酶Ya基因异源生物应答元件（XRE）具有组成型和异源生物诱导活性。我们目前的证据表明，XRE受本构C / EBP转录因子和异源生物激活的二恶英受体的调节。通过特异性抗体免疫耗竭和高纯度受体的结合，表明配体激活的XRE结合蛋白是二恶英受体。通过与C / EBP结合位点竞争，蛋白-DNA交联以确定组成蛋白的分子量，特异性抗体免疫耗竭和纯化的结合，证明了C / EBPα为组成型结合蛋白的鉴定。细菌表达的C / EBPα。 XRE的突变分析表明，组成型因子（C / EBP alpha）与二恶英受体具有几乎相同的重叠结合位点。在推定的C / EBP-XRE相互作用的功能测试中，共转染的C / EBP alpha在非异源生物应答的HeLa细胞系中激活了XRE测试启动子。出乎意料的是，共转染的C / EBP alpha对基础活性没有影响，但显着增加了异种生物应答的C / EBP阳性HepG2细胞系中XRE测试启动子的异种生物应答。此外，通过转染C / EBP寡核苷酸抑制HepG2细胞中的C / EBP结合蛋白抑制了异种应答。这些结果表明，C / EBPα和二恶英受体识别相同的DNA序列元件，并且通过这两个转录因子之间的协同相互作用可以发生转录调控。

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