Cui Zhicheng, Esposito Alessandra, Napolitano Gennaro, Ballabio Andrea, Hurley James H
Department of Molecular and Cell Biology, University of California Berkeley; Berkeley CA 94720, USA.
California Institute for Quantitative Biosciences, University of California, Berkeley, CA, 94720, USA.
bioRxiv. 2024 Nov 15:2024.11.15.623810. doi: 10.1101/2024.11.15.623810.
Mechanistic target of rapamycin complex 1 (mTORC1), which consists of mTOR, Raptor, and mLST8, receives signaling inputs from growth factor signals and nutrients. These signals are mediated by the Rheb and Rag small GTPases, respectively, which activate mTORC1 on the cytosolic face of the lysosome membrane. We biochemically reconstituted the activation of mTORC1 on membranes by physiological submicromolar concentrations of Rheb, Rags, and Ragulator. We determined the cryo-EM structure and found that Raptor and mTOR directly interact with the membrane at anchor points separated by up to 230 Å across the membrane surface. Full engagement of the membrane anchors is required for maximal activation, which is brought about by alignment of the catalytic residues in the mTOR kinase active site. The observations show at the molecular and atomic scale how converging signals from growth factors and nutrients drive mTORC1 recruitment to and activation on the lysosomal membrane in a three-step process, consisting of (1) Rag-Ragulator-driven recruitment to within ~100 Å of the lysosomal membrane, (2) Rheb-driven recruitment to within ~40 Å, and finally (3) direct engagement of mTOR and Raptor with the membrane. The combination of Rheb and membrane engagement leads to full catalytic activation, providing a structural explanation for growth factor and nutrient signal integration at the lysosome.
雷帕霉素复合物1(mTORC1)的作用机制靶点由mTOR、Raptor和mLST8组成,可接收来自生长因子信号和营养物质的信号输入。这些信号分别由Rheb和Rag小GTP酶介导,它们在溶酶体膜的胞质面上激活mTORC1。我们通过生理亚微摩尔浓度的Rheb、Rags和Ragulator在膜上对mTORC1的激活进行了生化重构。我们确定了冷冻电镜结构,发现Raptor和mTOR在膜表面相隔高达230 Å的锚定点处直接与膜相互作用。膜锚的完全结合是最大激活所必需的,这是由mTOR激酶活性位点中催化残基的排列实现的。这些观察结果在分子和原子尺度上展示了来自生长因子和营养物质的汇聚信号如何通过一个三步过程驱动mTORC1募集到溶酶体膜并在其上激活,该过程包括:(1)Rag-Ragulator驱动的募集到溶酶体膜约100 Å范围内;(2)Rheb驱动的募集到约40 Å范围内;最后(3)mTOR和Raptor与膜的直接结合。Rheb与膜结合的组合导致完全的催化激活,为溶酶体处生长因子和营养信号整合提供了结构解释。