Cloning of a mu-class glutathione S-transferase gene and identification of the glucocorticoid regulatory domains in its 5' flanking sequence.
Cloning of a mu-class glutathione S-transferase gene and identification of the glucocorticoid regulatory domains in its 5' flanking sequence.
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mu 类谷胱甘肽 S-转移酶基因的克隆以及其 5 侧翼序列中糖皮质激素调节结构域的鉴定。
DOI：
10.1073/pnas.89.13.6104
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发表时间：
1992
影响因子：
11.1
通讯作者：
Norris,JS
中科院分区：
文献类型：
--
作者：
Fan,W;Trifiletti,R;Cooper,T;Norris,JS
从仓鼠平滑肌肿瘤细胞 (DDT1 MF-2) 中分离出的 mu 类谷胱甘肽 S-转移酶基因 (hGSTYBX) 的表达受到糖皮质激素的转录上调，并且这种激素调节依赖于蛋白质合成。为了研究调控机制，我们对 hGSTYBX 基因组 DNA（包括其 5' 侧翼区域）进行了克隆和测序。 hGSTYBX 基因包含分散在 6.3 千碱基区域的 9 个外显子。当与氯霉素乙酰转移酶 (CAT) 报告基因连接时，5' 侧翼区域能够指导报告基因的转录。通过 5' 缺失研究，我们将主要的糖皮质激素诱导调节元件定位在核苷酸 -353 和 -239 之间。在该区域内，没有鉴定出经典的糖皮质激素反应元件 (TGTTCT)，但四个潜在的螺旋-环-螺旋结合域嵌入两个 16 碱基对重复序列中。另一个糖皮质激素调节结构域位于核苷酸-239 和-136 之间。 Cycloheximide 阻断糖皮质激素诱导的 -353CAT 和 -239CAT 报告基因（分别与 CAT 报告基因连接的 hGSTYBX 的核苷酸 -447 至 -12 和核苷酸 -239 至 -12）的转录；因此，我们的观察结果支持先前的结果，表明糖皮质激素诱导 hGSTYBX 是继发反应。
The expression of a mu-class glutathione S-transferase gene (hGSTYBX) isolated from hamster smooth muscle tumor cells (DDT1 MF-2) is transcriptionally up-regulated by glucocorticoids, and this hormonal regulation is dependent upon protein synthesis. To study the mechanism of regulation, we have cloned and sequenced hGSTYBX genomic DNA including its 5' flanking region. The hGSTYBX gene contains nine exons dispersed over a 6.3-kilobase region. When linked to a chloramphenicol acetyltransferase (CAT) reporter gene, the 5' flanking region was able to direct transcription of the reporter gene. With 5' deletion studies, we have localized the major glucocorticoid-inducible regulatory element between nucleotides -353 and -239. Within this region no classic glucocorticoid response element (TGTTCT) was identified, but four potential helix-loop-helix binding domains are embedded in two 16-base-pair repeats. Another glucocorticoid regulatory domain has been localized between nucleotides -239 and -136. Cycloheximide blocks glucocorticoid-induced transcription of both the -353CAT and -239CAT reporter genes (nucleotides -447 to -12 and nucleotides -239 to -12 of hGSTYBX, respectively, ligated to a CAT reporter gene); therefore, our observations support previous results suggesting that hGSTYBX induction by glucocorticoids is a secondary response.