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Title: Structural basis of the atypical activation mechanism of KRASV14I

Abstract

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 KRASV14I, 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 KRASA146T, 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 KRASA146T and KRASV14I that may have implications for themore » regulation of these variants and for the development of therapeutic strategies to manage KRAS variant-associated disorders.« less

Authors:
 [1];  [1];  [2];  [1];  [1]; ORCiD logo [1]; ORCiD logo [2]; ORCiD logo [1]
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  1. Univ. of Texas Southwestern Medical Center, Dallas, TX (United States)
  2. Northeastern Univ., Boston, MA (United States)
Publication Date:
Research Org.:
Argonne National Lab. (ANL), Argonne, IL (United States). Advanced Photon Source (APS)
Sponsoring Org.:
USDOE; National Institutes of Health (NIH); USDOD
OSTI Identifier:
1570737
Grant/Contract Number:  
U54CA196519; W81XWH-16-1-0106
Resource Type:
Accepted Manuscript
Journal Name:
Journal of Biological Chemistry
Additional Journal Information:
Journal Volume: 294; Journal Issue: 38; Journal ID: ISSN 0021-9258
Publisher:
American Society for Biochemistry and Molecular Biology
Country of Publication:
United States
Language:
ENGLISH
Subject:
59 BASIC BIOLOGICAL SCIENCES; 37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY; ras protein; GTPase; GTPase Kras (KRAS); guanine nucleotide-exchange factor (GEF); x-ray crystallography; Noonan syndrome; nucleotide exchange; RAS; RASopathy

Citation Formats

Bera, Asim K., Lu, Jia, Wales, Thomas E., Gondi, Sudershan, Gurbani, Deepak, Nelson, Andrew, Engen, John R., and Westover, Kenneth D. Structural basis of the atypical activation mechanism of KRASV14I. United States: N. p., 2019. Web. doi:10.1074/jbc.ra119.009131.
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Bera, Asim K., Lu, Jia, Wales, Thomas E., Gondi, Sudershan, Gurbani, Deepak, Nelson, Andrew, Engen, John R., & Westover, Kenneth D. Structural basis of the atypical activation mechanism of KRASV14I. United States. https://doi.org/10.1074/jbc.ra119.009131
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Bera, Asim K., Lu, Jia, Wales, Thomas E., Gondi, Sudershan, Gurbani, Deepak, Nelson, Andrew, Engen, John R., and Westover, Kenneth D. Wed . "Structural basis of the atypical activation mechanism of KRASV14I". United States. https://doi.org/10.1074/jbc.ra119.009131. https://www.osti.gov/servlets/purl/1570737.
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@article{osti_1570737,
title = {Structural basis of the atypical activation mechanism of KRASV14I},
author = {Bera, Asim K. and Lu, Jia and Wales, Thomas E. and Gondi, Sudershan and Gurbani, Deepak and Nelson, Andrew and Engen, John R. and Westover, Kenneth D.},
abstractNote = {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 KRASV14I, 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 KRASA146T, 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 KRASA146T and KRASV14I that may have implications for the regulation of these variants and for the development of therapeutic strategies to manage KRAS variant-associated disorders.},
doi = {10.1074/jbc.ra119.009131},
journal = {Journal of Biological Chemistry},
number = 38,
volume = 294,
place = {United States},
year = {Wed Jul 24 00:00:00 EDT 2019},
month = {Wed Jul 24 00:00:00 EDT 2019}
}
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https://doi.org/10.1074/jbc.ra119.009131
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