INSULIN STIMULATES GLYCERALDEHYDE-3-PHOSPHATE DEHYDROGENASE GENE-EXPRESSION THROUGH CIS-ACTING DNA-SEQUENCES
INSULIN STIMULATES GLYCERALDEHYDE-3-PHOSPHATE DEHYDROGENASE GENE-EXPRESSION THROUGH CIS-ACTING DNA-SEQUENCES
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DOI：
10.1073/pnas.85.14.5092
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发表时间：
1988-07-01
影响因子：
11.1
通讯作者：
RAMAIKA, C
中科院分区：
文献类型：
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作者：
ALEXANDER, MC;LOMANTO, M;RAMAIKA, C
甘油醛-3-磷酸脱氢酶[GAPDH; D-甘油醛-3-磷酸：NAD+氧化还原酶（磷酸化），EC 1.2.1.12] mRNA水平在培养的3 T3-F442 A脂肪细胞和H35肝癌细胞系中由生理浓度的胰岛素诱导。为了研究胰岛素调节这两种胰岛素敏感组织中GADPH mRNA水平的机制，我们分离了一个功能性的人GAPDH基因。当在3 T3-F442 A前脂肪细胞和H35肝癌细胞中稳定转染和表达时，完整的人GAPDH基因在3 T3-F442 A脂肪细胞中被胰岛素诱导10倍，在H35肝癌细胞系中被胰岛素诱导3倍，这与用内源基因获得的诱导相似。含有与氯霉素乙酰转移酶基因融合的人基因的序列-487至+20的人GAPDH-氯霉素乙酰转移酶构建体在稳定转染的3 T3脂肪细胞和稳定或瞬时转染的H35肝癌细胞系中受胰岛素调节，而劳斯肉瘤病毒与氯霉素乙酰转移酶融合蛋白则不受胰岛素调节。因此，胰岛素对人GAPDH基因表达的诱导作用是通过位于人GAPDH基因-487和+20之间的顺式作用序列介导的。
Glyceraldehyde-3-phosphate dehydrogenase [GAPDH; D-glyceraldehyde-3-phosphate:NAD+ oxidoreductase (phosphorylating), EC 1.2.1.12] mRNA levels are induced by physiologic concentrations of insulin in cultured 3T3-F442A adipocyte and H35 hepatoma cell lines. To examine the mechanism by which insulin regulates GADPH mRNA levels in these two insulin-sensitive tissues, we have isolated a functional human GAPDH gene. When stably transfected and expressed in 3T3-F442A preadipocytes and H35 hepatoma cells, the intact human GAPDH gene is induced 10-fold by insulin in 3T3-F442A adipocytes and 3-fold by insulin in H35 hepatoma lines, which is similar to the induction obtained with the endogenous gene. A human GAPDH-chloramphenicol acetyltrasnferase construct, containing sequnces -487 to +20 of the human gene fused to the chloramphenicol acetyltransferase gene, is regulated by insulin in stably transfected 3T3 adipocytes and stably or transiently transfected H35 hepatoma cell lines, whereas the Rous sarcoma virus.sbd.chloramphenicol acetyltransferase fusion protein is not. Thus, the inductive effect of insulin on human GAPDH gene expression is mediated through cis-acting sequences located between -487 and +20 of the human GAPDH gene.