Identification of the first mammalian sphingosine phosphate lyase gene and its functional expression in yeast.
Identification of the first mammalian sphingosine phosphate lyase gene and its functional expression in yeast.
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DOI：
10.1006/bbrc.1997.7993
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发表时间：
1998-01
影响因子：
3.1
通讯作者：
Jianhui Zhou;J. Saba
中科院分区：
文献类型：
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作者：
Jianhui Zhou;J. Saba
1-磷酸鞘氨醇 (S-1-P) 已被证明参与哺乳动物细胞的增殖信号转导途径。 1-磷酸鞘氨醇裂解酶 (SPL) 催化 S-1-P 的分解。使用线虫 SPL 核苷酸序列，我们鉴定了小鼠 EST 作为编码该酶的同源基因的推定候选者。小鼠 EST 测序揭示了 1707 个核苷酸的开放阅读框。该假定的小鼠 SPL 基因与线虫 SPL 基因有 62% 相似性和 39% 相同性，与酵母 SPL 基因有 59% 同源性和 39.6% 相同性。小鼠 SPL 基因在酵母菌株 delta bst1 中表达，该菌株携带 SPL 基因缺失并对鞘氨醇过敏，恢复了鞘氨醇抗性表型，表明该小鼠基因在表达时可以在功能上补充酵母缺陷。使用这些抗鞘氨醇转化体的提取物进行的体外酶测定证实了该小鼠 cDNA 克隆编码的 SPL 活性。 Northern分析表明小鼠SPL基因在不同组织中以不同水平表达。染色体定位将该 SPL 基因定位到 10 号染色体 32 cM 处。在这里，我们报告了第一个哺乳动物磷酸鞘氨醇裂解酶基因的鉴定。
Sphingosine-1-phosphate (S-1-P) has been shown to participate in the proliferative signal transduction pathways of mammalian cells. Sphingosine-1-phosphate lyase (SPL) catalyzes the breakdown of S-1-P. Using the C. elegans SPL nucleotide sequence, we identified a mouse EST as a putative candidate for the homologous gene encoding this enzyme. Sequencing of the mouse EST revealed an open reading frame of 1707 nucleotides. This putative mouse SPL gene is 62% similar and 39% identical to the C. elegans SPL gene and 59% homologous and 39.6% identical to the yeast SPL gene. Expression of the mouse SPL gene in a yeast strain-delta bst1, which carries a deletion of the SPL gene and is hypersensitive to sphingosine, restored a sphingosine-resistant phenotype, suggesting this mouse gene can functionally complement the yeast defect when expressed. In vitro enzyme assay using extracts from these sphingosine-resistant transformants confirmed the SPL activities encoded by this mouse cDNA clone. Northern analysis indicated the mouse SPL gene is expressed at various levels in different tissues. Chromosomal localization mapped this SPL gene to Chromosome 10 at 32 cM. Here, we report the identification of the first mammalian sphingosine phosphate lyase gene.