Cystic fibrosis transmembrane regulator regulates uptake of sphingoid base phosphates and lysophosphatidic acid - Modulation of cellular activity of spingosine 1-phosphate
Cystic fibrosis transmembrane regulator regulates uptake of sphingoid base phosphates and lysophosphatidic acid - Modulation of cellular activity of spingosine 1-phosphate
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DOI：
10.1074/jbc.m105442200
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发表时间：
2001-09-21
影响因子：
4.8
通讯作者：
Obeid, LM
中科院分区：
文献类型：
--
作者：
Boujaoude, LC;Bradshaw-Wilder, C;Obeid, LM
鞘脂与细胞生长、分化和程序性细胞死亡的调节有关。1-磷酸鞘氨醇（SPP）是近年来发现的一种重要的脂质信使和内皮分化基因受体蛋白家族的配体，并通过其介导其生物学效应。本实验室最近在酿酒酵母中的研究表明，ATP结合盒蛋白家族的一员酵母寡霉素抗性基因（YOR 1）参与了SPP的转运。囊性纤维化跨膜调节因子是ATP结合盒转运蛋白家族的独特成员，与YOR 1具有高度同源性。因此，我们着手调查，如果这个家庭的成员可以调节SPP运输。我们证明，C127/囊性纤维化跨膜调节（CFTR）细胞，表达野生型CFTR，表现出显着更高的摄取鞘氨醇1-磷酸比表达突变CFTR C127/Δ F508或C127/模拟转染细胞的细胞。这种作用是特异性的、剂量依赖性的，并被1-磷酸二氢鞘氨醇和溶血磷脂酸竞争。在不同细胞系中，鞘氨醇、C16-神经酰胺、鞘磷脂、溶血鞘磷脂、磷脂酰胆碱、溶血磷脂酰胆碱或磷脂酸的摄取无差异。用毛喉素或异丁基甲基黄嘌呤刺激cAMP的预处理不影响任何细胞系的摄取。此外，我们还发现，与C127/mock转染的细胞相比，C127/CFTR中SPP对丝裂原活化蛋白激酶的活化反应较低，这表明CFTR对SPP的摄取可能会使其偏离与细胞表面受体的相互作用并减弱信号传导功能。总之，这些数据表明CFTR在SPP和相关磷酸化脂质二氢鞘氨醇1-磷酸和溶血磷脂酸的摄取中。这种摄取影响SPP通过内皮分化基因受体调节生物活性的可用性。这些研究可能对囊性纤维化有重要意义。
Sphingolipids have been implicated in the regulation of cell growth, differentiation, and programmed cell death. Sphingosine 1-phosphate (SPP) has recently emerged as an important lipid messenger and a ligand for the endothelial differentiation gene receptor family of proteins through which it mediates its biologic effects. Recent studies in Saccharomyces cerevisiae in our laboratory implicated the yeast oligomycin resistance gene (YOR1), a member of the ATP binding cassette family of proteins, in the transport of SPP. The cystic fibrosis transmembrane regulator is a unique member of the ATP binding cassette transporter family and has high homology with YOR1. We therefore set out to investigate if this member of the family can regulate SPP transport. We demonstrate that C127/cystic fibrosis transmembrane regulator (CFTR) cells, expressing wild type CFTR, exhibited significantly higher uptake of sphingosine 1-phosphate than either cells expressing a mutant CFTR C127/Delta F508 or C127/mock-transfected cells. This effect was specific, dose-dependent, and competed off by dihydrosphingosine 1-phosphate and lysophosphatidic acid. There was no difference in uptake of sphingosine, C16-ceramide, sphingomyelin, lysophingomyelin, phosphatidylcholine, lysophosphatidylcholine, or phosphatidic acid among the different cell lines. Pretreatment with forskolin or isobutylmethylxanthine to stimulate cAMP did not affect the uptake in any of the cell lines. Moreover, we found that mitogen-activated protein kinase activation by SPP was less responsive in C127/CFTR as compared with C127/mock-transfected cells, suggesting that uptake of SPP by CFTR may divert it from interacting with its cell surface receptors and attenuate signaling functions. Taken together, these data implicate CFTR in uptake of SPP and the related phosphorylated lipids dihydrosphingosine 1-phosphate and lysophosphatidic acid. This uptake influences the availability of SPP to modulate biologic activity via endothelial differentiation gene receptors. These studies may have important implications to cystic fibrosis.