Differential antagonism of Ras biological activity by catalytic and Src homology domains of Ras GTPase activation protein

Geoffrey J. Clark, Lawrence Quilliam, Mark M. Hisaka, Channing J. Der

Research output: Contribution to journalArticle

53 Citations (Scopus)

Abstract

Ras p120 GTPase activation protein (GAP), a cytosolic protein, is a negative mediator and potential downstream effector of Ras function. Since membrane association is critical for Ras function, we introduced the Ras membrane-targeting signal (a 19-residue peptide ending in CAAX, where C = cysteine, A = aliphatic amino acid, and X = any amino acid) onto the GAP N-terminal Src homology 2 and 3 and the C-terminal catalytic domains (designated nGAP/CAAX and cGAP/CAAX, respectively) to determine the role of membrane association in GAP function. cGAP/CAAX and full-length GAP/CAAX, but not GAP or nGAP/CAAX, exhibited potent growth inhibitory activity. Whereas both oncogenic and normal Ras activity were inhibited by cGAP/CAAX, nGAP/CAAX, despite lacking the Ras binding domain, inhibited the activity of oncogenic Ras without affecting the action of normal Ras. Altogether, these results demonstrate that membrane association potentiates GAP catalytic activity, support an effector function for GAP, and suggest that normal and oncogenic Ras possess different downstream interactions.

Original languageEnglish (US)
Pages (from-to)4887-4891
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume90
Issue number11
StatePublished - Jun 1 1993
Externally publishedYes

Fingerprint

ras Proteins
src Homology Domains
GTP Phosphohydrolases
Proteins
Membranes
Amino Acids
Cysteine
Catalytic Domain
Fatty Acids
Peptides
Growth

Keywords

  • CAAX motif
  • Transactivation
  • Transformation

ASJC Scopus subject areas

  • General
  • Genetics

Cite this

Differential antagonism of Ras biological activity by catalytic and Src homology domains of Ras GTPase activation protein. / Clark, Geoffrey J.; Quilliam, Lawrence; Hisaka, Mark M.; Der, Channing J.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 90, No. 11, 01.06.1993, p. 4887-4891.

Research output: Contribution to journalArticle

@article{78225d158df74af68868efb90b71efdb,
title = "Differential antagonism of Ras biological activity by catalytic and Src homology domains of Ras GTPase activation protein",
abstract = "Ras p120 GTPase activation protein (GAP), a cytosolic protein, is a negative mediator and potential downstream effector of Ras function. Since membrane association is critical for Ras function, we introduced the Ras membrane-targeting signal (a 19-residue peptide ending in CAAX, where C = cysteine, A = aliphatic amino acid, and X = any amino acid) onto the GAP N-terminal Src homology 2 and 3 and the C-terminal catalytic domains (designated nGAP/CAAX and cGAP/CAAX, respectively) to determine the role of membrane association in GAP function. cGAP/CAAX and full-length GAP/CAAX, but not GAP or nGAP/CAAX, exhibited potent growth inhibitory activity. Whereas both oncogenic and normal Ras activity were inhibited by cGAP/CAAX, nGAP/CAAX, despite lacking the Ras binding domain, inhibited the activity of oncogenic Ras without affecting the action of normal Ras. Altogether, these results demonstrate that membrane association potentiates GAP catalytic activity, support an effector function for GAP, and suggest that normal and oncogenic Ras possess different downstream interactions.",
keywords = "CAAX motif, Transactivation, Transformation",
author = "Clark, {Geoffrey J.} and Lawrence Quilliam and Hisaka, {Mark M.} and Der, {Channing J.}",
year = "1993",
month = "6",
day = "1",
language = "English (US)",
volume = "90",
pages = "4887--4891",
journal = "Proceedings of the National Academy of Sciences of the United States of America",
issn = "0027-8424",
number = "11",

}

TY - JOUR

T1 - Differential antagonism of Ras biological activity by catalytic and Src homology domains of Ras GTPase activation protein

AU - Clark, Geoffrey J.

AU - Quilliam, Lawrence

AU - Hisaka, Mark M.

AU - Der, Channing J.

PY - 1993/6/1

Y1 - 1993/6/1

N2 - Ras p120 GTPase activation protein (GAP), a cytosolic protein, is a negative mediator and potential downstream effector of Ras function. Since membrane association is critical for Ras function, we introduced the Ras membrane-targeting signal (a 19-residue peptide ending in CAAX, where C = cysteine, A = aliphatic amino acid, and X = any amino acid) onto the GAP N-terminal Src homology 2 and 3 and the C-terminal catalytic domains (designated nGAP/CAAX and cGAP/CAAX, respectively) to determine the role of membrane association in GAP function. cGAP/CAAX and full-length GAP/CAAX, but not GAP or nGAP/CAAX, exhibited potent growth inhibitory activity. Whereas both oncogenic and normal Ras activity were inhibited by cGAP/CAAX, nGAP/CAAX, despite lacking the Ras binding domain, inhibited the activity of oncogenic Ras without affecting the action of normal Ras. Altogether, these results demonstrate that membrane association potentiates GAP catalytic activity, support an effector function for GAP, and suggest that normal and oncogenic Ras possess different downstream interactions.

AB - Ras p120 GTPase activation protein (GAP), a cytosolic protein, is a negative mediator and potential downstream effector of Ras function. Since membrane association is critical for Ras function, we introduced the Ras membrane-targeting signal (a 19-residue peptide ending in CAAX, where C = cysteine, A = aliphatic amino acid, and X = any amino acid) onto the GAP N-terminal Src homology 2 and 3 and the C-terminal catalytic domains (designated nGAP/CAAX and cGAP/CAAX, respectively) to determine the role of membrane association in GAP function. cGAP/CAAX and full-length GAP/CAAX, but not GAP or nGAP/CAAX, exhibited potent growth inhibitory activity. Whereas both oncogenic and normal Ras activity were inhibited by cGAP/CAAX, nGAP/CAAX, despite lacking the Ras binding domain, inhibited the activity of oncogenic Ras without affecting the action of normal Ras. Altogether, these results demonstrate that membrane association potentiates GAP catalytic activity, support an effector function for GAP, and suggest that normal and oncogenic Ras possess different downstream interactions.

KW - CAAX motif

KW - Transactivation

KW - Transformation

UR - http://www.scopus.com/inward/record.url?scp=0027279039&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0027279039&partnerID=8YFLogxK

M3 - Article

C2 - 8506332

AN - SCOPUS:0027279039

VL - 90

SP - 4887

EP - 4891

JO - Proceedings of the National Academy of Sciences of the United States of America

JF - Proceedings of the National Academy of Sciences of the United States of America

SN - 0027-8424

IS - 11

ER -