Comprehensive identification of PIP3-regulated PH domains from C elegans to H sapiens by model prediction and live imaging
Comprehensive identification of PIP3-regulated PH domains from C elegans to H sapiens by model prediction and live imaging
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DOI：
10.1016/j.molcel.2008.04.008
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发表时间：
2008-05-09
期刊：
影响因子：
16
通讯作者：
Teruel, Mary N.
中科院分区：
文献类型：
--
作者：
Park, Wei Sun;Do Heo, Won;Teruel, Mary N.
磷酸肌醇3-激酶（PI 3 K）及其产物磷脂酰肌醇（3，4，5）-三磷酸（PIP 3）通过将具有普列克底物蛋白同源性（PH）结构域和可能的其它结构域的蛋白募集到质膜（PM）来控制细胞生长、迁移和其它过程。然而，以前的实验和结构工作与PH结构域留下了相互矛盾的证据，哪些是PIP 3调节。在这里，我们使用130个YFP缀合的小鼠PH结构域的活细胞共聚焦成像，发现20%响应受体产生的PIP 3产生而转移到PM。我们开发了一种递归学习算法来预测来自不同真核生物的1200个PH结构域的PIP 3调节，并验证了它准确地预测PIP 3调节。引人注目的是，该算法表明，PIP 3调控是由PH结构域上的氨基酸指定的，而不仅仅是PIP 3结合口袋，并且必须独立于PIP 3不敏感的祖先PH结构域进化多次。最后，我们的算法和活细胞实验提供了不同物种中PH结构域的功能调查，表明PI 3 K调节从大约两个C. elegans和4种Drosophiia到40种脊椎动物蛋白质。
Phosphoinositide 3-kinase (PI3K) and its product phosphatidylinositol(3,4,5)-trisphosphate (PIP3) control cell growth, migration, and other processes by recruiting proteins with pleckstrin homology (PH) domains and possibly other domains to the plasma membrane (PM). However, previous experimental and structural work with PH domains left conflicting evidence about which ones are PIP3 regulated. Here we used live-cell confocal imaging of 130 YFP-conjugated mouse PH domains and found that 20% translocated to the PM in response to receptor-generated PIP3 production. We developed a recursive-learning algorithm to predict PIP3 regulation of 1200 PH domains from different eukaryotes and validated that it accurately predicts PIP3 regulation. Strikingly, this algorithm showed that PIP3 regulation is specified by amino acids across the PH domain, not just the PIP3-binding pocket, and must have evolved several times independently from PIP3-insensitive ancestral PH domains. Finally, our algorithm and live-cell experiments provide a functional survey of PH domains in different species, showing that PI3K regulation increased from approximately two C. elegans and four Drosophiia to 40 vertebrate proteins.