Structural basis of the atypical activation mechanism of KRASV14I

Bera, Asim K.; Lu, Jia; Wales, Thomas E.; Gondi, Sudershan; Gurbani, Deepak; Nelson, Andrew; Engen, John R.; Westover, Kenneth D.

doi:10.1074/jbc.ra119.009131

Title: Structural basis of the atypical activation mechanism of KRAS^V14I

Journal Article · 23 July 2019 · Journal of Biological Chemistry

DOI: https://doi.org/10.1074/jbc.ra119.009131 · OSTI ID:1570737

Bera, Asim K. ^[1]; Lu, Jia ^[1]; Wales, Thomas E. ^[2]; Gondi, Sudershan ^[1]; Gurbani, Deepak ^[1];

^[1];

^[2];

^[1]

Univ. of Texas Southwestern Medical Center, Dallas, TX (United States)
Northeastern Univ., Boston, MA (United States)

RAS regulation and signaling are largely accomplished by direct protein-protein interactions, making RAS protein dynamics a critical determinant of RAS function. In this study, we report a crystal structure of GDP-bound KRAS^V14I, a mutated KRAS variant associated with the developmental RASopathy disorder Noonan syndrome (NS), at 1.5–1.6 Å resolution. The structure is notable for revealing a marked extension of switch 1 away from the G-domain and nucleotide-binding site of the KRAS protein. We found that this extension is associated with a loss of the magnesium ion and a tilt in the position of the guanine base because of the additional carbon introduced by the isoleucine substitution. Hydrogen-deuterium exchange MS analysis confirmed that this conformation occurs in solution, but also disclosed a difference in kinetics when compared with KRAS^A146T, another RAS mutant that displays a nearly identical conformation in previously reported crystal structures. This conformational change contributed to a high rate of guanine nucleotide-exchange factor (GEF)-dependent and -independent nucleotide exchange and to an increase in affinity for SOS Ras/Rac GEF 1 (SOS1), which appears to be the major mode of activation for this RAS variant. These results highlight a mechanistic connection between KRAS^A146T and KRAS^V14I that may have implications for the regulation of these variants and for the development of therapeutic strategies to manage KRAS variant-associated disorders.

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Research Organization:: Argonne National Lab. (ANL), Argonne, IL (United States). Advanced Photon Source (APS)

Sponsoring Organization:: National Institutes of Health (NIH); USDOD; USDOE

OSTI ID:: 1570737

Journal Information:: Journal of Biological Chemistry, Journal Name: Journal of Biological Chemistry Journal Issue: 38 Vol. 294; ISSN 0021-9258

Publisher:: American Society for Biochemistry and Molecular BiologyCopyright Statement

Country of Publication:: United States

Language:: ENGLISH

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59 BASIC BIOLOGICAL SCIENCES
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Title: Structural basis of the atypical activation mechanism of KRASV14I

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Title: Structural basis of the atypical activation mechanism of KRAS^V14I