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Spatial organization of PI3K-PI(3,4,5)P3-AKT signaling by focal adhesions

Mol Cell. 2024-11; 
Jing Wang , Zhengyang An , Zhongsheng Wu , Wei Zhou , Pengyu Sun , Piyu Wu , Song Dang , Rui Xue , Xue Bai , Yongtao Du , Rongmei Chen , Wenxu Wang , Pei Huang , Sin Man Lam , Youwei Ai , Suling Liu , Guanghou Shui , Zhe Zhang , Zheng Liu , Jianyong Huang , Xiaohong Fang , Kangmin He
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5X Sample Buffer Cells were solubilized at 4°C for 30 min in RIPA lysis buffer (50 mM Tris-HCl, pH 8.0, 150?mM NaCl, 1% NP-40, 0.5% sodium deoxycholate, 0.1% SDS) with a protease and phosphatase inhibitor cocktail (Thermo Fisher Scientific, 78442) and then pelleted at 13,400 g for 15?min at 4°C. The supernatant was mixed with 5× sample buffer (GenScript, MB01015), heated to 100°C for 10 min, and then fractionated by SDS–PAGE and transferred to nitrocellulose membranes (Pall, 66485). Get A Quote

摘要

The class I phosphatidylinositol 3-kinase (PI3K)-AKT signaling pathway is a key regulator of cell survival, growth, and proliferation and is among the most frequently mutated pathways in cancer. However, where and how PI3K-AKT signaling is spatially activated and organized in mammalian cells remains poorly understood. Here, we identify focal adhesions (FAs) as subcellular signaling hubs organizing the activation of PI3K-PI(3,4,5)P3-AKT signaling in human cancer cells containing p110α mutations under basal conditions. We find that class IA PI3Ks are preferentially recruited to FAs for activation, resulting in localized production of PI(3,4,5)P3 around FAs. As the effector protein of PI(3,4,5)P3, AKT1 molecules ... More

關鍵詞

AKT; FAK; PI(3,4,5)P(3); class I PI3K; focal adhesion; lipid signaling; single-molecule imaging.
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