Kuwayama Hidekazu, Kikuchi Haruhisa, Oshima Yoshiteru, Kubohara Yuzuru
Faculty of Life and Environmental Sciences, University of Tsukuba, Tsukuba 305-8572, Japan.
Laboratory of Natural Product Chemistry, Graduate School of Pharmaceutical Sciences, Tohoku University, Sendai 980-8578, Japan.
Biochem Biophys Rep. 2016 Sep 19;8:219-226. doi: 10.1016/j.bbrep.2016.09.006. eCollection 2016 Dec.
In the development of the cellular slime mold , two chlorinated compounds, the differentiation-inducing factors DIF-1 and DIF-2, play important roles in the regulation of both cell differentiation and chemotactic cell movement. However, the receptors of DIFs and the components of DIF signaling systems have not previously been elucidated. To identify the receptors for DIF-1 and DIF-2, we here performed DIF-conjugated affinity gel chromatography and liquid chromatography-tandem mass spectrometry and identified the glutathione S-transferase GST4 as a major DIF-binding protein. Knockout and overexpression mutants of ( and , respectively) formed fruiting bodies, but the fruiting bodies of cells were smaller than those of wild-type Ax2 cells, and those of cells were larger than those of Ax2 cells. Both chemotaxis regulation and stalk cell formation by DIFs in the mutants were similar to those of Ax2 cells. These results suggest that GST4 is a DIF-binding protein that regulates the sizes of cell aggregates and fruiting bodies in .
在细胞黏菌的发育过程中，两种氯化化合物，即分化诱导因子DIF-1和DIF-2，在细胞分化和趋化性细胞运动的调节中发挥着重要作用。然而，DIFs的受体和DIF信号系统的组成成分此前尚未阐明。为了鉴定DIF-1和DIF-2的受体，我们在此进行了DIF偶联亲和凝胶色谱和液相色谱-串联质谱分析，并鉴定出谷胱甘肽S-转移酶GST4为主要的DIF结合蛋白。（分别为 和 的）基因敲除和过表达突变体形成了子实体，但 细胞的子实体比野生型Ax2细胞的小，而 细胞的子实体比Ax2细胞的大。在 突变体中，DIFs对趋化性的调节和柄细胞的形成与Ax2细胞相似。这些结果表明，GST4是一种DIF结合蛋白，可调节 中细胞聚集体和子实体的大小。
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