NUCLEOTIDE-SEQUENCE ANALYSIS OF MURINE 21-HYDROXYLASE GENES - MUTATIONS AFFECTING GENE-EXPRESSION
NUCLEOTIDE-SEQUENCE ANALYSIS OF MURINE 21-HYDROXYLASE GENES - MUTATIONS AFFECTING GENE-EXPRESSION
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DOI：
10.1073/pnas.83.24.9601
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发表时间：
1986-12-01
影响因子：
11.1
通讯作者：
PARKER, KL
中科院分区：
文献类型：
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作者：
CHAPLIN, DD;GALBRAITH, LJ;PARKER, KL
类固醇21-羟化酶[21- oase；类固醇21-monooxygenase;类固醇，氢供体：氧氧化还原酶（21-羟基化）；EC 1.14.99.10]是一种细胞色素P-450酶，是肾上腺合成矿物皮质激素和糖皮质激素所必需的。编码该蛋白的基因存在于小鼠主要组织相容性复合体S区的两个拷贝（21- oase AS和B）中。先前利用基因特异性寡核苷酸探针和基因转移的研究表明，只有21- oase A基因在BALB/c小鼠中表达。在这里，我们展示了两个BALB/c 21- oase编码基因的完整初级结构。核苷酸序列的比较确定了跨越21- oase B基因第二外显子的215个核苷酸的缺失；21- oase B基因的其他核苷酸变化引入了框架移位和过早终止的约束子。C57BL/6和DBA/2J小鼠的Southern blot分析表明，这些菌株中存在类似的缺失；然而，C3H/HeJ菌株是一种结构变体。将21- oase B启动子置于21- oase A结构序列的5‘’位置组成的杂交基因转染到Y1肾上腺皮质肿瘤细胞后有效转录。这些发现表明21- oase B基因启动子是功能性的，并提示21- oase B结构基因的突变是其缺乏表达的原因。
Steroid 21-hydroxylase [21-OHase; steroid 21-monooxygenase; steroid, hydrogen-donor:oxygen oxidoreductase (21-hydroxylating); EC 1.14.99.10] is a cytochrome P-450 enzyme required for the adrenal synthesis of mineralocorticoids and glucocorticoids. The gene encoding this protein is present in two copies (21-OHase AS and B) in the S region of the murine major histocompatibility complex. Previous studies utilizing gene-specific oligonucleotide probes and gene transfer showed that only the 21-OHase A gene is expressed in the BALB/c mouse. Here, we present the complete primary structures of both BALB/c 21-OHase encoding genes. Comparison of the nucleotide sequences defines a deletion of 215 nucleotides spanning the second exon of the 21-OHase B gene; other nucleotide changes in the 21-OHase B gene introduce frame shifts and premature termination condons. Southern blot analysis of C57BL/6 and DBA/2J mice indicates that a similar deletion is present in these strains; however the C3H/HeJ strain is a structural variant. A hybrid gene composed of the 21-OHase B promoter placed 5'' of the 21-OHase A structural sequences was efficiently transcribed following transfection into Y1 adrenocortical tumor cells. These findings demonstrate that the 21-OHase B gene promoter is functional and suggest that mutations within the 21-OHase B structural gene are responsible for its lack of expression.