Chemical genetic analysis of FTY720‐ and Ca2+‐sensitive mutants reveals a functional connection between FTY720 and membrane trafficking
Chemical genetic analysis of FTY720‐ and Ca2+‐sensitive mutants reveals a functional connection between FTY720 and membrane trafficking
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FTY720 和 Ca2+ 敏感突变体的化学遗传分析揭示了 FTY720 与膜运输之间的功能联系
DOI：
10.1111/gtc.12800
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发表时间：
2020
期刊：
影响因子：
2.1
通讯作者：
Sugiura Reiko
中科院分区：
文献类型：
--
作者：
Hagihara Kanako;Kanda Yuki;Ishida Kouki;Satoh Ryosuke;Takasaki Teruaki;Maeda Takuya;Sugiura Reiko
FTY 720是一种鞘氨醇-1-磷酸（S1 P）类似物，用作治疗多发性硬化症的免疫调节剂。越来越多的证据表明，FTY 720的作用模式不依赖于S1 P调节剂。在裂殖酵母中，FTY 720诱导细胞内Ca 2+和ROS水平增加。我们以前已经确定了49个基因，其中缺失导致FTY 720敏感性。在这里，我们根据FTY 720敏感突变体与Ca 2+稳态的相关性来表征FTY 720敏感突变体，并鉴定了16个FTY 720和Ca 2+敏感突变体（fcs突变体）。大多数FTY 720敏感突变体显示出升高的Ca 2+水平，并通过FTY 720处理表现出Ca 2+失调。在其缺失使得细胞对FTY 720和Ca 2+敏感的基因中的功能类别之一包括高尔基体/内体膜运输。值得注意的是，FTY 720，而不是不能诱导Ca 2+增加的磷酸化FTY 720，抑制了野生型细胞中酸性磷酸酶的分泌。重要的是，分泌缺陷的高尔基体/内体运输突变体，Vps 45，或Ryh 1缺失，进一步加剧FTY 720。我们的突变体筛选还鉴定了腺苷酸环化酶相关蛋白Cap 1和Rictor同源物Ste 20，其缺失显著加剧了FTY 720敏感性分泌障碍。总的来说，我们的数据可能表明FTY 720与分泌干扰相结合对增殖和Ca 2+稳态的协同影响。
FTY720, a sphingosine‐1‐phosphate (S1P) analog, is used as an immune modulator to treat multiple sclerosis. Accumulating evidence has suggested the mode of action of FTY720 independent of an S1P modulator. In fission yeast, FTY720 induces an increase in intracellular Ca2+and ROS levels. We have previously identified 49 genes of which deletion causes FTY720 sensitivity. Here, we characterized the FTY720‐sensitive mutants in terms of their relevance to the Ca2+homeostasis and identified the 16FTY720‐ andCa2+‐sensitive mutants (fcsmutants). Most of the FTY720‐sensitive mutants showed elevated Ca2+levels and exhibited Ca2+dysregulation by FTY720 treatment. One of the functional categories among the genes whose deletion renders cells susceptible to FTY720 and Ca2+include the Golgi/endosomal membrane trafficking. Notably, FTY720, but not phosphorylated FTY720 incapable of inducing Ca2+increase, inhibited the secretion of acid phosphatase in the wild‐type cells. Importantly, secretory defects of the Golgi/endosomal trafficking mutants, Vps45, or Ryh1 deletion, were further exacerbated by FTY720. Ourfcsmutant screen also identified the adenylyl cyclase‐associated protein Cap1 and a Rictor homolog Ste20, whose deletion markedly exacerbated FTY720‐sensitive secretory impairment. Collectively, our data may suggest a synergistic impact of FTY720 combined with secretion perturbation on proliferation and Ca2+homeostasis.