The Dictyostelium prestalk inducer differentiation-inducing factor-1 (DIF-1) triggers unexpectedly complex global phosphorylation changes.
The Dictyostelium prestalk inducer differentiation-inducing factor-1 (DIF-1) triggers unexpectedly complex global phosphorylation changes.
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DOI：
10.1091/mbc.e14-08-1319
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发表时间：
2015-02-15
影响因子：
3.3
通讯作者：
Williams JG
中科院分区：
文献类型：
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作者：
Sugden C;Urbaniak MD;Araki T;Williams JG
DIF-1 是一种聚酮化合物，主要被定性为转录调节因子，但对改变 DIF 处理细胞中磷酸化水平的肽进行蛋白质组范围的鉴定揭示了许多新的靶点。因此，基因转录的小分子调节剂作为蛋白质修饰的多功能调节剂发挥着额外的作用。分化诱导因子-1 (DIF-1) 是一种聚酮化合物，可诱导阿米巴盘基网柄菌分化为前茎细胞。我们使用三标记 SILAC 方法对添加 DIF-1 后最初几分钟内发生的磷酸化变化进行了全局定量筛选。这揭示了 DIF-1 控制的信号传导的新世界，其中包括 MAPK 和蛋白激酶 B 信号传导途径的成分、放线肌球蛋白细胞骨架信号传导网络的成分以及广泛的小 GTP 酶及其调节因子的变化。结果还提供证据表明 Ca2+/钙调蛋白依赖性磷酸酶钙调神经磷酸酶在 DimB 前茎转录因子的 DIF-1 信号传导中发挥作用。在全球水平上，DIF-1 导致磷酸化/去磷酸化平衡向净去磷酸化方向发生重大转变。有趣的是，许多响应 DIF-1 去磷酸化的位点在响应细胞外 cAMP 信号传导时被磷酸化。这与表明两种诱导剂之间存在拮抗作用的研究一致，也与我们观察到的响应 DIF-1 的 cAMP 受体快速去磷酸化以及已知的 DIF-1 对 cAMP 趋化性的抑制作用一致。所有 MS 数据均可通过 ProteomeXchange 获得，标识符为 PXD001555。
DIF-1, a polyketide, has been primarily characterized as a transcriptional regulator, but proteome-wide identification of peptides that change phosphorylation level in DIF-treated cell reveals many novel targets. Thus a small-molecule regulator of gene transcription plays additional roles as a multifunctional regulator of protein modification. Differentiation-inducing factor-1 (DIF-1) is a polyketide that induces Dictyostelium amoebae to differentiate as prestalk cells. We performed a global quantitative screen for phosphorylation changes that occur within the first minutes after addition of DIF-1, using a triple-label SILAC approach. This revealed a new world of DIF-1–controlled signaling, with changes in components of the MAPK and protein kinase B signaling pathways, components of the actinomyosin cytoskeletal signaling networks, and a broad range of small GTPases and their regulators. The results also provide evidence that the Ca2+/calmodulin–dependent phosphatase calcineurin plays a role in DIF-1 signaling to the DimB prestalk transcription factor. At the global level, DIF-1 causes a major shift in the phosphorylation/dephosphorylation equilibrium toward net dephosphorylation. Of interest, many of the sites that are dephosphorylated in response to DIF-1 are phosphorylated in response to extracellular cAMP signaling. This accords with studies that suggest an antagonism between the two inducers and also with the rapid dephosphorylation of the cAMP receptor that we observe in response to DIF-1 and with the known inhibitory effect of DIF-1 on chemotaxis to cAMP. All MS data are available via ProteomeXchange with identifier PXD001555.