EXPRESSION OF MURINE 21-HYDROXYLASE IN MOUSE ADRENAL-GLANDS AND IN TRANSFECTED Y1 ADRENOCORTICAL TUMOR-CELLS
EXPRESSION OF MURINE 21-HYDROXYLASE IN MOUSE ADRENAL-GLANDS AND IN TRANSFECTED Y1 ADRENOCORTICAL TUMOR-CELLS
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DOI：
10.1073/pnas.82.23.7860
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发表时间：
1985-01-01
影响因子：
11.1
通讯作者：
SCHIMMER, BP
中科院分区：
文献类型：
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作者：
PARKER, KL;CHAPLIN, DD;SCHIMMER, BP
小鼠主要组织相容性复合体的S区含有两个结构上相关的基因(21-OHase A和21-OHase B)，它们编码21-羟基酶(21-OHase)，21-OHase是肾上腺类固醇合成所必需的酶。通过使用针对21-OHase A和B基因的寡核苷酸探针和DNA介导的基因转移来分析这两个基因的表达。寡核苷酸与BALB/c肾上腺RNA斑点杂交表明，所有的21-OHase基因都来自21-OHase A基因。携带21-OHase A或B基因的Cosmids通过与pSV2-neo共切割的方法导入Y1肾上腺肿瘤细胞。类固醇代谢实验和RNA印迹杂交结果显示，转21-OHase A基因的细胞表达21-OHase，亲本Y1细胞和转21-OHase B基因的细胞均未检测到21-OHase的转录本。用促肾上腺皮质激素处理21-OHase A转基因细胞后，21-OHase mRNA水平增加了10倍，从而模拟了这种激素对原代肾上腺培养中21-OHase水平的影响。21-OHase A基因在Y1细胞中的调节表达为研究肾上腺特异性调节21-OHase活性的因素提供了一个有用的系统。
The S region of the murine major histocompatibility complex contains two structurally related genes (21-OHase A and 21-OHase B) that encode 21-hydroxylase (21-OHase), an enzyme essential for the synthesis of adrenal steriods. Expression of these two genes has been analyzed by using oligonucleotide probes specific for the 21-OHase A and B genes and by DNA-mediated gene transfer. Hybridization of the oligonucleotides to blots of BALB/c adrenal RNA demonstrated that all 21-OHase mRNA is derived from the 21-OHase A gene. Cosmids bearing either the 21-OHase A or B gene were introduced into Y1 adrenocortical tumor cells by cotransection with pSV2-neo. Cells transfected with the 21-OHase A gene expressed 21-OHase as determined by steroid metabolism and by RNA blot hybridization; 21-OHase transcripts were not detected in parent Y1 cells or in cells transfected with the 21-OHase B gene. Treatment of 21-OHase A transfectants with adrenocorticotropin increased 21-OHase mRNA levels by up to 10-fold, thus mimicking the observed effect of this hormone on 21-OHase levels in primary adrenal cultures. The regulated expression of the 21-OHase A gene in transfected Y1 cells should provide a useful system for the investigation of factors controlling the adrenal-specific regulation of 21-OHase activity.