Phenotypic Characterization of a Comprehensive Set of MAPK1/ERK2 Missense Mutants

Tumor-specific genomic information has the potential to guide therapeutic strategies and revolutionize patient treatment. Currently, this approach is limited by an abundance of disease-associated mutants whose biological functions and impacts on therapeutic response are uncharacterized. To begin to address this limitation, we functionally characterized nearly all (99.84%) missense mutants of MAPK1/ERK2, an essential effector of oncogenic RAS and RAF. Using this approach, we discovered rare gain- and loss-of-function ERK2 mutants found in human tumors, revealing that, in the context of this assay, mutational frequency alone cannot identify all functionally impactful mutants. Gain-of-function ERK2 mutants induced variable responses to RAF-, MEK-, and ERK-directed therapies, providing a reference for future treatment decisions. Tumor-associated mutations spatially clustered in two ERK2 effector-recruitment domains yet produced mutants with opposite phenotypes. This approach articulates an allele-characterization framework that can be scaled to meet the goals of genome-guided oncology. [Display omitted] •Characterization of function and ERK-inhibitor sensitivity of 6,810 ERK2 mutants•Discovery of rare tumor-associated gain- and loss-of-function ERK2 mutants•A mechanistic class of ERK2 GOF mutants that mimic the sevenmaker mutation is defined•Mutation of distinct ERK2 effector domains leads to opposite activity profiles Using comprehensive functional characterization of ERK2 mutants, Brenan et al. identify rare cancer-associated gain- and loss-of-function mutant ERK2 proteins. Gain-of-function ERK2 mutants form two mechanistic classes that drive differential responses to RAF, MEK, and ERK inhibitors, which may help to indicate therapeutic treatment strategies for patients whose tumors harbor these mutants.