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首页> 外文期刊>Cell biology international. >Exportin 1-independent nuclear export of GAPDH.
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Exportin 1-independent nuclear export of GAPDH.

机译：GAPDH的Exportin 1独立核出口。

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摘要

Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is a key enzyme of the glycolytic pathway. Recent studies have demonstrated an additional role in apoptosis: GAPDH is targeted to the nucleus during apoptotic signalling. This nuclear transport has also been observed in serum-depleted cells, but it is reversible in fibroblasts, in contrast to apoptotic-induced transport (Eur J Cell Biol 80 (2001) 419). Here, we analyse the serum depletion-induced transport processes of GAPDH in NIH 3T3 cells. Prolonged serum depletion did not cause cell death, nuclear fragmentation (hoechst staining) or a significant increase in DNA strand-breaks (comet assay). Using cells expressing green fluorescent protein (GFP)-tagged GAPDH allowed us to monitor its intracellular localisation by confocal laser scanning microscopy (CLSM). Treatment of cells with the exportin1 inhibitor leptomycin B (LMB) did not influence cytoplasmic localisation of GFP-GAPDH, indicating that nuclear targeting of GAPDH is not constitutive and may be altered via a serum-dependent regulatory export process. Suprisingly, the export of nuclear GFP-GAPDH after re-addition of serum to starved cells was not prevented by LMB. Thus, nuclear export of GAPDH upon serum depletion is not mediated by exportin1.
机译：3-磷酸​​甘油醛脱氢酶（GAPDH）是糖酵解途径的关键酶。最近的研究表明在细胞凋亡中还有其他作用：GAPDH在细胞凋亡信号转导过程中靶向细胞核。还已经在贫血的细胞中观察到了这种核转运，但是与凋亡诱导的转运相反，它在成纤维细胞中是可逆的（Eur J Cell Biol 80（2001）419）。在这里，我们分析了血清耗竭诱导的GAPDH在NIH 3T3细胞中的转运过程。长时间的血清耗竭不会导致细胞死亡，核分裂（hoechst染色）或DNA链断裂显着增加（彗星分析）。使用表达绿色荧光蛋白（GFP）标签的GAPDH的细胞使我们能够通过共聚焦激光扫描显微镜（CLSM）监测其细胞内定位。用exportin1抑制剂瘦霉素B（LMB）处理细胞不会影响GFP-GAPDH的细胞质定位，这表明GAPDH的核靶向作用不是组成性的，并且可能会通过血清依赖性调节性输出过程而改变。出乎意料的是，LMB不能阻止将血清重新添加到饥饿细胞后核GFP-GAPDH的输出。因此，血清枯竭后GAPDH的核输出不是由exportin1介导的。

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