Kong Kyoung-Ah, Park Sungdo, Park Hyoungwoo, Kim Myoung Hee
Department of Anatomy, Embryology Laboratory, Brain Korea 21 Project for Medical Sciences, Yonsei University College of Medicine, Seoul 120-752, Korea.
Ann N Y Acad Sci. 2003 Dec;1010:433-6. doi: 10.1196/annals.1299.078.
A novel gene Jpk (Jopock) has been originally isolated through yeast 1 hybridization technique as a trans-acting factor interacting with the position-specific regulatory element of a murine Hoxa-7. Northern analysis revealed that the Jpk was expressed at day 7.0 post coitum (p.c.) during early gastrulation. Previously it has been shown that a trace amount of JPK protein led bacterial cells to death. In eukaryotic F9 cells, Jpk also led the cell to death-generating DNA ladder: fewer than 50% of the cells survived after 72-h transfection. Flow cytometric analysis with cells stained with each Annexin V/7-amino-actinomycin D (7-AAD), MitoTracker, and hydroethidine (HE) revealed that Jpk induced apoptotic cell death in a time-dependent manner, reduced mitochondrial membrane potential, and increased ROS (reactive oxygen species) production, respectively. Additionally, Jpk seemed to regulate the Bcl family at the transcriptional level when RT-PCR was performed. Although the precise mechanism is not clear, these results altogether suggest that Jpk is a potent inducer of apoptosis through generation of ROS as well as concomitant reduction of mitochondrial membrane potential.
一种新基因Jpk（Jopock）最初是通过酵母单杂交技术作为与小鼠Hoxa - 7的位置特异性调控元件相互作用的反式作用因子分离出来的。Northern分析显示，Jpk在早期原肠胚形成阶段的妊娠第7.0天（p.c.）表达。此前已表明，微量的JPK蛋白会导致细菌细胞死亡。在真核F9细胞中，Jpk也会导致细胞产生DNA梯状条带从而死亡：转染72小时后，存活的细胞不到50%。用膜联蛋白V/7-氨基放线菌素D（7-AAD）、线粒体追踪染料和氢乙啶（HE）对细胞进行染色后的流式细胞术分析表明，Jpk分别以时间依赖性方式诱导凋亡细胞死亡、降低线粒体膜电位并增加活性氧（ROS）的产生。此外，进行RT-PCR时，Jpk似乎在转录水平上调节Bcl家族。尽管确切机制尚不清楚，但这些结果共同表明，Jpk是通过产生ROS以及伴随线粒体膜电位降低而诱导凋亡的强效诱导剂。
