Universität zu Köln

Yin, Guowei ORCID: 0000-0003-2669-7160, Kistler, Samantha, George, Samuel D., Kuhlmann, Nora, Garvey, Leslie, Bagni, Rachel K., Lammers, Michael ORCID: 0000-0003-4168-4640, Der, Channing J. and Campbell, Sharon L. (2017). A KRAS GTPase K104Q Mutant Retains Downstream Signaling by Offsetting Defects in Regulation. J. Biol. Chem., 292 (11). S. 4446 - 4457. BETHESDA: AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC. ISSN 1083-351X

Full text not available from this repository.

Abstract

The KRAS GTPase plays a critical role in the control of cellular growth. The activity of KRAS is regulated by guanine nucleotide exchange factors (GEFs), GTPase-activating proteins (GAPs), and also post-translational modification. Lysine 104 in KRAScan be modified by ubiquitylation and acetylation, but the role of this residue in intrinsic KRAS function has not been well characterized. We find that lysine 104 is important for GEF recognition, because mutations at this position impaired GEF-mediated nucleotide exchange. Because the KRAS K104Q mutant has recently been employed as an acetylation mimetic, we conducted a series of studies to evaluate its in vitro and cellbased properties. Herein, we found that KRAS K104Q exhibited defects in both GEF-mediated exchange and GAP-mediated GTP hydrolysis, consistent with NMR-detected structural perturbations in localized regions of KRAS important for recognition of these regulatory proteins. Despite the partial defect in both GEF and GAP regulation, KRAS K104Q did not alter steady-state GTP-bound levels or the ability of the oncogenic KRAS G12V mutant to cause morphologic transformation of NIH 3T3 mouse fibroblasts and of WT KRAS to rescue the growth defect of mouse embryonic fibroblasts deficient in all Ras genes. We conclude that the KRAS K104Q mutant retains bothWTand mutant KRAS function, probably due to offsetting defects in recognition of factors that up-regulate (GEF) and down-regulate (GAP) RAS activity.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Yin, Guowei UNSPECIFIED orcid.org/0000-0003-2669-7160 UNSPECIFIED Kistler, Samantha UNSPECIFIED UNSPECIFIED UNSPECIFIED George, Samuel D. UNSPECIFIED UNSPECIFIED UNSPECIFIED Kuhlmann, Nora UNSPECIFIED UNSPECIFIED UNSPECIFIED Garvey, Leslie UNSPECIFIED UNSPECIFIED UNSPECIFIED Bagni, Rachel K. UNSPECIFIED UNSPECIFIED UNSPECIFIED Lammers, Michael UNSPECIFIED orcid.org/0000-0003-4168-4640 UNSPECIFIED Der, Channing J. UNSPECIFIED UNSPECIFIED UNSPECIFIED Campbell, Sharon L. UNSPECIFIED UNSPECIFIED UNSPECIFIED
URN:	urn:nbn:de:hbz:38-236699
DOI:	10.1074/jbc.M116.762435
Journal or Publication Title:	J. Biol. Chem.
Volume:	292
Number:	11
Page Range:	S. 4446 - 4457
Date:	2017
Publisher:	AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC
Place of Publication:	BETHESDA
ISSN:	1083-351X
Language:	English
Faculty:	Unspecified
Divisions:	Unspecified
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language GUANINE-NUCLEOTIDE EXCHANGE; NUCLEAR-MAGNETIC-RESONANCE; RAS ACTIVATOR SON; K-RAS; BINDING PROTEINS; STRUCTURAL BASIS; H-RAS; ACETYLATION; EFFECTORS; DYNAMICS Multiple languages Biochemistry & Molecular Biology Multiple languages
Refereed:	Yes
URI:	http://kups.ub.uni-koeln.de/id/eprint/23669