Structure-function analysis of the SHOC2-MRAS-PP1C holophosphatase complex.
Structure-function analysis of the SHOC2-MRAS-PP1C holophosphatase complex.
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DOI：
10.1038/s41586-022-04928-2
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发表时间：
2022-09
期刊：
影响因子：
64.8
通讯作者：
Aguirre, Andrew J.
中科院分区：
文献类型：
--
作者：
Kwon, Jason J.;Hajian, Behnoush;Bian, Yuemin;Young, Lucy C.;Amor, Alvaro J.;Fuller, James R.;Fraley, Cara V.;Sykes, Abbey M.;So, Jonathan;Pan, Joshua;Baker, Laura;Lee, Sun Joo;Wheeler, Douglas B.;Mayhew, David L.;Persky, Nicole S.;Yang, Xiaoping;Root, David E.;Barsotti, Anthony M.;Stamford, Andrew W.;Perry, Charles K.;Burgin, Alex;McCormick, Frank;Lemke, Christopher T.;Hahn, William C.;Aguirre, Andrew J.
受体酪氨酸激酶（RTK）-RAS信号通过下游促分裂原活化蛋白激酶（MAPK）级联调节细胞增殖和存活。SHOC 2-MRAS-PP 1C全磷酸酶复合物通过去除RAF家族蛋白上的抑制性磷酸化以增强MAPK信号传导而作为RTK-RAS信号传导的关键调节剂发挥功能。SHOC 2与MRAS和PP 1C形成三元复合物，该复合物的人种系功能获得性突变导致先天性RAS病综合征。然而，这种复合物的结构和组装知之甚少。在这里，我们使用低温电子显微镜（cryo-EM）来解析SHOC 2-MRAS-PP 1C复合物的结构，分辨率为2.9 nm。此外，我们定义了全酶相互作用的生物物理学原理，阐明了复合物的组装顺序，并通过深度突变扫描系统地询问了SHOC 2几乎所有可能的错义变体的功能后果。我们证明SHOC 2通过富含亮氨酸的重复区域的凹面结合PP 1C和MRAS，并通过含有隐蔽的RNF基序的N-末端无序区域进一步接合PP 1C。初始复合物的形成由SHOC 2-PP 1C相互作用介导，并通过与负载GTP的MRAS结合而稳定。这些观察定义了RASopathies和癌症中SHOC 2突变体如何稳定复合体成员的相互作用以增强全磷酸酶活性。总之，这种整合的结构-功能模型全面定义了SHOC 2-MRAS-PP 1C全磷酸酶复合物内的关键结合相互作用，以告知治疗开发。
Receptor tyrosine kinase (RTK)-RAS signaling through the downstream mitogen activated protein kinase (MAPK) cascade regulates cell proliferation and survival. The SHOC2-MRAS-PP1C holophosphatase complex functions as a key regulator of RTK-RAS signaling by removing an inhibitory phosphorylation on RAF family proteins to potentiate MAPK signaling. SHOC2 forms a ternary complex with MRAS and PP1C, and human germline gain-of-function mutations of this complex result in congenital RASopathy syndromes. However, the structure and assembly of this complex are poorly understood. Here, we use cryogenic electron microscopy (cryo-EM) to resolve the structure of the SHOC2-MRAS-PP1C complex to 2.9Å resolution. Furthermore, we define the biophysical principles of holoenzyme interactions, elucidate the assembly order of the complex, and systematically interrogate the functional consequence of nearly all possible missense variants of SHOC2 through deep mutational scanning. We demonstrate that SHOC2 binds PP1C and MRAS through the concave surface of the leucine-rich repeat region and further engages PP1C through the N-terminal disordered region containing a cryptic RVxF motif. Initial complex formation is mediated by SHOC2-PP1C interactions and is stabilized by binding of GTP-loaded MRAS. These observations define how SHOC2 mutants in RASopathies and cancer stabilize interactions of complex members to enhance holophosphatase activity. Together, this integrative structure-function model comprehensively defines key binding interactions within the SHOC2-MRAS-PP1C holophosphatase complex to inform therapeutic development.
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影响因子：
48
作者：
Barad BA;Echols N;Wang RY;Cheng Y;DiMaio F;Adams PD;Fraser JS
通讯作者：
Fraser JS
影响因子：
3.7
作者：
Beard H;Cholleti A;Pearlman D;Sherman W;Loving KA
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Loving KA
影响因子：
30.8
作者：
Cordeddu, Viviana;Di Schiavi, Elia;Pennacchio, Len A.;Ma'ayan, Avi;Sarkozy, Anna;Fodale, Valentina;Cecchetti, Serena;Cardinale, Alessio;Martin, Joel;Schackwitz, Wendy;Lipzen, Anna;Zampino, Giuseppe;Mazzanti, Laura;Digilio, Maria C.;Martinelli, Simone;Flex, Elisabetta;Lepri, Francesca;Bartholdi, Deborah;Kutsche, Kerstin;Ferrero, Giovanni B.;Anichini, Cecilia;Selicorni, Angelo;Rossi, Cesare;Tenconi, Romano;Zenker, Martin;Merlo, Daniela;Dallapiccola, Bruno;Iyengar, Ravi;Bazzicalupo, Paolo;Gelb, Bruce D.;Tartaglia, Marco
通讯作者：
Tartaglia, Marco
DOI：
10.1042/bcj20210708
发表时间：
2021-12-22
期刊：
The Biochemical journal
影响因子：
--
作者：
Kimanius D;Dong L;Sharov G;Nakane T;Scheres SHW
通讯作者：
Scheres SHW
DOI：
10.1107/s2059798318006551
发表时间：
2018-06-01
期刊：
Acta crystallographica. Section D, Structural biology
影响因子：
--
作者：
Afonine PV;Poon BK;Read RJ;Sobolev OV;Terwilliger TC;Urzhumtsev A;Adams PD
通讯作者：
Adams PD