Role of Ad4-binding protein/steroidogenic factor 1 in regulating NADPH production in adrenocortical Y-1 cells
Role of Ad4-binding protein/steroidogenic factor 1 in regulating NADPH production in adrenocortical Y-1 cells
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DOI：
10.1507/endocrj.ej16-0467
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发表时间：
2017-01-01
影响因子：
2
通讯作者：
Morohashi, Ken-ichirou
中科院分区：
文献类型：
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作者：
Li, Bing;Baba, Takashi;Morohashi, Ken-ichirou
Ad 4结合蛋白/类固醇生成因子1（Ad 4 BP/SF-1）是核受体超家族的成员，在类固醇生成细胞中表达并调节所有类固醇生成基因的表达。我们最近采用mRNA和染色质免疫沉淀序列（ChIP-seq）来证明Ad 4 BP/SF-1直接调节几乎所有糖酵解基因的表达。戊糖磷酸途径（PPP）有助于烟酰胺腺嘌呤二核苷酸磷酸（NADPH）的产生。Ad 4 BP/SF-1基因敲低后，PPP基因和细胞内NADPH的表达均降低，但这些基因并不是Ad 4 BP/SF-1的直接靶基因。因此，本研究调查Ad 4 BP/SF-1是否直接调节参与NADPH产生的基因。对先前发表的mRNA序列（mRNA-seq）和ChIP-seq数据集的检查强烈表明，其他NADPH产生基因，如苹果酸酶1（Me 1）和亚甲基四氢叶酸脱氢酶2（Mthfd 2），可能是Ad 4 BP/SF-1的直接靶点。报告基因测定和细胞内NADPH浓度的测定支持Ad 4 BP/SF-1通过调节这些基因来调节NADPH产生的观点。NADPH是化合物如类固醇的大分子合成和活性氧的解毒所必需的。当合成类固醇激素时，类固醇生成细胞通过类固醇生成P450介导的酶促反应消耗NADPH。NADPH还通过消除作为P450反应的副产物产生的活性氧物质而被消耗。总体而言，Ad 4 BP/SF-1可能通过参与消耗和供应的生物过程的基因的协同调节来维持细胞内NADPH水平。
Ad4-binding protein/steroidogenic factor 1 (Ad4BP/SF-1), a member of the nuclear receptor superfamily, is expressed in steroidogenic cells and regulates all steroidogenic gene expression. We recently employed mRNA and chromatin immunoprecipitation sequence (ChIP-seq) to demonstrate that Ad4BP/SF-1 directly regulates the expression of nearly all glycolytic genes. The pentose phosphate pathway (PPP) contributes to the production of nicotinamide adenine dinucleotide phosphate (NADPH). Although the expression of PPP genes and intracellular NADPH were decreased by Ad4BP/SF-1 knockdown, these genes were not the direct targets of Ad4BP/SF-1. This study therefore investigates whether Ad4BP/SF-1 directly regulates genes implicated in NADPH production. Examination of previously published data sets of mRNA sequence (mRNA-seq) and ChIP-seq strongly suggested a possibility that other NADPH-producing genes, such as malic enzyme 1 (Me1) and methylenetetrahydrofolate dehydrogenase 2 (Mthfd2), are the direct targets of Ad4BP/SF-1. Reporter gene assays and determination of intracellular NADPH concentration supported the notion that Ad4BP/SF-1 regulates NADPH production by regulating these genes. NADPH is required for macromolecule synthesis of compounds such as steroids, and for detoxification of reactive oxygen species. When synthesizing steroid hormones, steroidogenic cells consume NADPH through enzymatic reactions mediated by steroidogenic P450s. NADPH is also consumed through elimination of reactive oxygen species produced as the byproducts of the P450 reactions. Overall, Ad4BP/SF-1 potentially maintains the intracellular NADPH level through cooperative regulation of genes involved in the biological processes for consumption and supply.