ZNF143 mediates basal and tissue-specific expression of human transaldolase
ZNF143 mediates basal and tissue-specific expression of human transaldolase
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DOI：
10.1074/jbc.m307039200
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发表时间：
2004-03-26
影响因子：
4.8
通讯作者：
Perl, A
中科院分区：
文献类型：
--
作者：
Grossman, CE;Qian, YM;Perl, A
转醛醇酶通过磷酸戊糖途径控制NADPH和5-磷酸核糖的产生来调节氧化还原依赖性细胞凋亡。足以驱动氯霉素乙酰转移酶报告基因活性的最小启动子被映射到相对于人转醛醇酶基因的转录起始位点的核苷酸-49至-1。用转醛醇酶表达细胞系的核提取物进行DNA酶I足迹分析揭示了核苷酸-29至-16的保护作用。电泳迁移率变动分析确定了一个单一的显性DNA-蛋白质复合物，该复合物被转录因子ZNF 143/76的共有序列或转醛醇酶启动子内ZNF 76/143基序的突变所废除。AP-2 α识别序列的突变，部分重叠ZNF 143基序，增加了HeLa细胞中的TAL-H启动子活性，而对不表达AP-2 α的HepG 2细胞没有显著影响。ZNF 143与AP-2 α的协同作用得到了HeLa细胞超移位分析的支持，其中AP-2可能作为TAL启动子活性的细胞类型特异性阻遏物。然而，在HepG 2和HeLa细胞中，全长ZNF 143、ZNF 76或ZNF 143的显性负性DNA结合结构域的过表达分别增强、维持或消除了转醛醇酶启动子的活性，表明ZNF 143从转醛醇酶核心启动子启动转录。ZNF 143过表达也增加了转醛醇酶的活性。ZNF 143和转醛醇酶的表达在21种不同的人体组织中相关，并且在哺乳期乳腺中与非哺乳期乳腺相比分别协同上调14倍和34倍。染色质免疫沉淀研究证实，ZNF 143/73与转醛醇酶启动子在体内。因此，ZNF 143在转醛醇酶的基础和组织特异性表达以及控制细胞存活和分化的代谢网络的调节中起关键作用。
Transaldolase regulates redox-dependent apoptosis through controlling NADPH and ribose 5-phosphate production via the pentose phosphate pathway. The minimal promoter sufficient to drive chloramphenicol acetyltransferase reporter gene activity was mapped to nucleotides - 49 to - 1 relative to the transcription start site of the human transaldolase gene. DNase I footprinting with nuclear extracts of transaldolase-expressing cell lines unveiled protection of nucleotides - 29 to - 16. Electrophoretic mobility shift assays identified a single dominant DNA-protein complex that was abolished by consensus sequence for transcription factor ZNF143/76 or mutation of the ZNF76/143 motif within the transaldolase promoter. Mutation of an AP-2alpha recognition sequence, partially overlapping the ZNF143 motif, increased TAL-H promoter activity in HeLa cells, without significant impact on HepG2 cells, which do not express AP-2alpha. Cooperativity of ZNF143 with AP-2alpha was supported by supershift analysis of HeLa cells where AP-2 may act as cell type-specific repressor of TAL promoter activity. However, overexpression of full-length ZNF143, ZNF76, or dominant-negative DNA-binding domain of ZNF143 enhanced, maintained, or abolished transaldolase promoter activity, respectively, in HepG2 and HeLa cells, suggesting that ZNF143 initiates transcription from the transaldolase core promoter. ZNF143 overexpression also increased transaldolase enzyme activity. ZNF143 and transaldolase expression correlated in 21 different human tissues and were coordinately upregulated 14- and 34-fold, respectively, in lactating mammary glands compared with nonlactating ones. Chromatin immunoprecipitation studies confirm that ZNF143/73 associates with the transaldolase promoter in vivo. Thus, ZNF143 plays a key role in basal and tissue-specific expression of transaldolase and regulation of the metabolic network controlling cell survival and differentiation.