A cross-talk between RNA splicing and signaling pathway alters Fas gene expression at post-transcriptional level: Alternative splicing of Fas mRNA in the leukemic U937 cells
A cross-talk between RNA splicing and signaling pathway alters Fas gene expression at post-transcriptional level: Alternative splicing of Fas mRNA in the leukemic U937 cells
复制标题
DOI：
10.1016/j.lab.2005.05.004
复制
发表时间：
2005-09-01
期刊：
影响因子：
--
通讯作者：
Eguchi, K
中科院分区：
文献类型：
--
作者：
Aratake, K;Kamachi, M;Eguchi, K
现在人们普遍认为，选择性剪接是一种以低遗传成本产生蛋白质复杂性的机制。然而，人们对控制关键凋亡调节因子选择性剪接的分子机制知之甚少。在这里，我们通过逆转录聚合酶链式反应 (RT-PCR) 研究促凋亡刺激对 Fas mRNA 选择性剪接的影响。将 U937 细胞暴露于依托泊苷、星形孢菌素、紫杉醇或环己酰胺促进了剪接变体的出现，从而保留了 152 个碱基对的内含子 5。用花萼素 A（一种蛋白磷酸酶-1 (PP-1) 抑制剂）和伏马菌素 B1（一种神经酰胺合成酶抑制剂）进行预处理，可防止依托泊苷诱导的选择性剪接Fas mRNA。我们的数据表明，RNA 剪接和信号通路之间通过内源性神经酰胺合成和随后的磷酸酶激活的串扰是一种在转录后水平修饰 Fas 基因表达的机制。
It is now widely accepted that alternative splicing is a mechanism that is responsible for generating protein complexity at low genetic cost. However, little is known about molecular mechanisms that govern alternative splicing of key apoptotic regulators. Here we investigate the effect of pro-apoptotic stimuli on alternative splicing of Fas mRNA by means of reverse transcription-polymerase chain reaction (RT-PCR). Exposure of U937 cells to etoposide, staurosporine, pacritaxel, or cyclohexamide promoted the appearance of the splice variant, which retained the 152-base-pair intron 5. Pretreatment with calyculin A, an inhibitor of protein phosphatase-1 (PP-1) as well as fumonisin B1, an inhibitor of ceramide synthase, prevented etoposide-induced alternative splicing of Fas mRNA. Our data demonstrate that cross-talk between RNA splicing and signaling pathways through endogenous ceramide synthesis and subsequent phosphatase activation is a mechanism that modifies Fas gene expression at the post-transcriptional level.