Structure of the G60A mutant of Ras: implications for the dominant negative effect

Bradley Ford; Karlheinz Skowronek; Sean Boykevisch; Dafna Bar-Sagi; Nicolas Nassar

doi:10.1074/jbc.M502240200

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Structure of the G60A mutant of Ras: implications for the dominant negative effect

Journal article

Open access

Peer reviewed

Structure of the G60A mutant of Ras: implications for the dominant negative effect

Bradley Ford, Karlheinz Skowronek, Sean Boykevisch, Dafna Bar-Sagi and Nicolas Nassar

The Journal of biological chemistry, Vol.280(27), pp.25697-25705

07/08/2005

DOI: 10.1074/jbc.M502240200

PMID: 15878843

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Abstract

Substituting alanine for glycine at position 60 in v-H-Ras generated a dominant negative mutant that completely abolished the ability of v-H-Ras to transform NIH 3T3 cells and to induce germinal vesicle breakdown in Xenopus oocytes. The crystal structure of the GppNp-bound form of RasG60A unexpectedly shows that the switch regions adopt an open conformation reminiscent of the structure of the nucleotide-free form of Ras in complex with Sos. Critical residues that normally stabilize the guanine nucleotide and the Mg(2+) ion have moved considerably. Sos binds to RasG60A but is unable to catalyze nucleotide exchange. Our data suggest that the dominant negative effect observed for RasG60A.GTP could result from the sequestering of Sos in a non-productive Ras-GTP-guanine nucleotide exchange factor ternary complex.

Glycine - genetics

Protein Structure, Tertiary

Xenopus

NIH 3T3 Cells

Chlorocebus aethiops

Protein Structure, Secondary

Guanosine Triphosphate - metabolism

Crystallography

Oncogene Protein p21(ras) - metabolism

Oocytes

Animals

Alanine - genetics

Oncogene Protein p21(ras) - chemistry

Protein Binding

Mice

Guanosine Diphosphate - metabolism

COS Cells

Amino Acid Substitution

Oncogene Protein p21(ras) - genetics

Details

Title: Subtitle: Structure of the G60A mutant of Ras: implications for the dominant negative effect
Creators: Bradley Ford - Department of Physiology and Biophysics, Stony Brook University, Stony Brook, New York 11794-8661, USA
Karlheinz Skowronek
Sean Boykevisch
Dafna Bar-Sagi
Nicolas Nassar
Resource Type: Journal article
Publication Details: The Journal of biological chemistry, Vol.280(27), pp.25697-25705
DOI: 10.1074/jbc.M502240200
PMID: 15878843
NLM abbreviation: J Biol Chem
ISSN: 0021-9258
eISSN: 1083-351X
Publisher: United States
Grant note: T32 GM008444 / NIGMS NIH HHS T32-DK07521-16 / NIDDK NIH HHS
Language: English
Date published: 07/08/2005
Academic Unit: Pathology
Record Identifier: 9984047724702771

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