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首页> 外文期刊>American Journal of Physiology >Synthesis of the Na-K-ATPase gamma-subunit is regulated at both the transcriptional and translational levels in IMCD3 cells.
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Synthesis of the Na-K-ATPase gamma-subunit is regulated at both the transcriptional and translational levels in IMCD3 cells.

机译：Na-K-ATPaseγ亚基的合成受IMCD3细胞转录和翻译水平的调节。

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摘要

We previously reported that hypertonicity-mediated upregulation of the gamma-subunit of Na-K-ATPase is dependent on both the JNK and the PI3 kinase pathways. The present experiments were undertaken to explore the mechanisms whereby these pathways regulate the expression of the gamma-subunit in inner medullary collecting duct cells (IMCD3). Inhibition of JNK with SP-600125 (20 muM), a concentration that causes an approximately 95% inhibition of hypertonicity-stimulated JNK activation, markedly decreased the amount of the gamma-subunit in response to 550 mosmol/kgH(2)O for 48 h. This was accompanied by a parallel decrease in the gamma-subunit mRNA. The rate at which the gamma-subunit mRNA decreased was unaffected by actinomycin D. In contrast, inhibition of PI3 kinase with LY-294002 results in a marked decrease in the amount of gamma-subunit protein but without alteration in gamma-subunit message. The rate at which the gamma-subunit protein decreased was unaffected by cyclohexamide. Transfection of IMCD3cells with a gamma-subunit construct results in the expression of both gamma-subunit message and protein. However, in cortical collecting duct cells (M1 cells) such transfection resulted in expression of only the message and not the protein. We conclude that JNK regulates the gamma-subunit at the transcriptional level while PI3 kinase regulates gamma-subunit expression at the translational level. There is also posttranscriptional cell specificity in the expression of the gamma -subunit of Na-K-ATPase.
机译：我们以前曾报道过，高渗介导的Na-K-ATPaseγ亚基上调依赖于JNK和PI3激酶途径。进行本实验以探索这些途径调节内髓收集管细胞（IMCD3）中γ亚基表达的机制。用SP-600125（20μM）抑制JNK的浓度会引起高渗刺激的JNK激活的约95％抑制，从而显着降低了550 mosmol / kgH（2）O响应48的γ亚基的量H。这伴随着γ-亚基mRNA的平行降低。 γ-亚基mRNA的降低速率不受放线菌素D的影响。相反，用LY-294002抑制PI3激酶可导致γ-亚基蛋白量显着减少，但不会改变γ-亚基信息。 γ-亚基蛋白减少的速率不受环己酰胺的影响。用γ-亚基构建体转染IMCD3细胞可导致γ-亚基信息和蛋白质的表达。但是，在皮层收集导管细胞（M1细胞）中，这种转染仅表达信息而不表达蛋白质。我们得出的结论是，JNK在转录水平上调节γ-亚基，而PI3激酶在翻译水平上调节γ-亚基的表达。 Na-K-ATP酶的γ-亚基的表达中也存在转录后细胞特异性。

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