DAPIT Over-Expression Modulates Glucose Metabolism and Cell Behaviour in HEK293T Cells.
DAPIT Over-Expression Modulates Glucose Metabolism and Cell Behaviour in HEK293T Cells.
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DOI：
10.1371/journal.pone.0131990
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Kainulainen H
中科院分区：
文献类型：
--
作者：
Kontro H;Cannino G;Rustin P;Dufour E;Kainulainen H
胰岛素敏感组织中的糖尿病相关蛋白（DAPIT）是线粒体ATP合成酶的一个亚基，也被发现与液泡H+-ATP酶有关。其表达在有氧代谢升高的细胞和积极运输营养物质和离子的上皮细胞中特别高。已知DAPIT的缺失会诱导线粒体ATP合酶的丧失，但其过表达的影响尚不清楚。为了研究DAPIT高表达的后果，我们构建了在人胚肾细胞HEK 293 T中组成型表达DAPIT的转基因细胞系。增强的DAPIT表达通过降低H+-ATP合酶活性而降低线粒体DNA含量和线粒体质量以及饱和呼吸链。DAPIT过表达还可增加线粒体膜电位和超氧化物水平，并将转录因子缺氧诱导因子1α（Hif 1 α）和β-catenin转运至细胞核。因此，与对照细胞相比，过表达DAPIT的细胞使用更多的葡萄糖并产生更大量的乳酸。有趣的是，这些变化与上皮间质（EMT）样的转变，改变E-钙粘蛋白N-钙粘蛋白和上调几个关键的连接/粘附蛋白。在生理水平，DAPIT过表达通过G1期阻滞和迁移减慢细胞生长，并增强细胞脱离。几种癌症还显示出Usmg 5（编码DAPIT的基因）的基因组拷贝数增加，从而为DAPIT可能在癌症中具有致癌功能提供了强有力的相关证据。因此，DAPIT过表达似乎调节线粒体功能并改变细胞调节，促进无氧代谢并诱导EMT样转变。我们认为DAPIT过表达将线粒体代谢的变化与生理和病理生理调节相结合，并表明它可能在H+-ATP合酶功能障碍中发挥关键作用。
Diabetes Associated Protein in Insulin-sensitive Tissues (DAPIT) is a subunit of mitochondrial ATP synthase and has also been found to associate with the vacuolar H+-ATPase. Its expression is particularly high in cells with elevated aerobic metabolism and in epithelial cells that actively transport nutrients and ions. Deletion of DAPIT is known to induce loss of mitochondrial ATP synthase but the effects of its over-expression are obscure. In order to study the consequences of high expression of DAPIT, we constructed a transgenic cell line that constitutively expressed DAPIT in human embryonal kidney cells, HEK293T. Enhanced DAPIT expression decreased mtDNA content and mitochondrial mass, and saturated respiratory chain by decreasing H+-ATP synthase activity. DAPIT over-expression also increased mitochondrial membrane potential and superoxide level, and translocated the transcription factors hypoxia inducible factor 1α (Hif1α) and β-catenin to the nucleus. Accordingly, cells over-expressing DAPIT used more glucose and generated a larger amount of lactate compared to control cells. Interestingly, these changes were associated with an epithelial to mesenchymal (EMT)-like transition by changing E-cadherin to N-cadherin and up-regulating several key junction/adhesion proteins. At physiological level, DAPIT over-expression slowed down cell growth by G1 arrest and migration, and enhanced cell detachment. Several cancers also showed an increase in genomic copy number of Usmg5 (gene encoding DAPIT), thereby providing strong correlative evidence for DAPIT possibly having oncogenic function in cancers. DAPIT over-expression thus appears to modulate mitochondrial functions and alter cellular regulations, promote anaerobic metabolism and induce EMT-like transition. We propose that DAPIT over-expression couples the changes in mitochondrial metabolism to physiological and pathophysiological regulations, and suggest it could play a critical role in H+-ATP synthase dysfunctions.
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影响因子：
24.5
作者：
Deng N;Goh LK;Wang H;Das K;Tao J;Tan IB;Zhang S;Lee M;Wu J;Lim KH;Lei Z;Goh G;Lim QY;Tan AL;Sin Poh DY;Riahi S;Bell S;Shi MM;Linnartz R;Zhu F;Yeoh KG;Toh HC;Yong WP;Cheong HC;Rha SY;Boussioutas A;Grabsch H;Rozen S;Tan P
通讯作者：
Tan P
影响因子：
64.8
作者：
通讯作者：
--
影响因子：
5
作者：
Benit, Paule;Goncalves, Sergio;Rustin, Pierre
通讯作者：
Rustin, Pierre
影响因子：
4.8
作者：
Cannino, Giuseppe;El-Khoury, Riyad;Dufour, Eric
通讯作者：
Dufour, Eric
影响因子：
3.5
作者：
Geromel, V;Kadhom, N;Rustin, P
通讯作者：
Rustin, P