Structure of the MRAS-SHOC2-PP1C phosphatase complex

RAS-MAPK signalling is fundamental for cell proliferation and is altered in most human cancers^1-3. However, our mechanistic understanding of how Ras signals through Raf is still incomplete. Although studies revealed snapshots for autoinhibited and active RAF-MEK1-14-3-3 complexes⁴, the intermediate steps that lead to Raf activation remain unclear. The MRAS-SHOC2-PP1C holophosphatase dephosphorylates Raf at serine 259, resulting in the partial displacement of 14-3-3 and RAF-RAS association^3,5,6. MRAS, SHOC2 and PP1C are mutated in rasopathies-developmental syndromes caused by aberrant MAPK pathway activation^6-14-and SHOC2 itself has emerged as potential target in receptor tyrosine kinase (RTK)-RAS-driven tumours^15-18. Despite its importance, structural understanding of the SHOC2 holophosphatase is lacking. Here we determine, using X-ray crystallography, the structure of the MRAS-SHOC2-PP1C complex. SHOC2 bridges PP1C and MRAS through its concave surface and enables reciprocal interactions between all three subunits. Biophysical characterization indicates a cooperative assembly driven by the MRAS GTP-bound active state, an observation that is extendible to Other RAS isoforms. Our findings support the concept of a RAS-driven and multi-molecular model for Raf activation in which individual RAS-GTP molecules recruit RAF-14-3-3 and SHOC2-PP1C to produce downstream pathway activation. Importantly, we find that rasopathy and Cancer mutations reside at protein-protein interfaces within the holophosphatase, resulting in enhanced affinities and function. Collectively, our findings shed light on a fundamental mechanism of Ras biology and on mechanisms of clinically observed enhanced RAS-MAPK signalling, therefore providing the structural basis for therapeutic interventions.