Sensitivity of wild type and mutant ras alleles to Ras specific exchange factors: Identification of factor specific requirements

Klaus Hvid Nielsen, Lars Gredsted, James Broach, Berthe Marie Willumsen

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

We have investigated the productive interaction between the four mammalian Ras proteins (H-, N-, KA- and KB-Ras) and their activators, the mammalian exchange factors mSos1, GRF1 and GRP, by using a modified Saccharomyces cerevisiae whose growth is dependent on activation of a mammalian Ras protein by its activator. All four mammalian Ras proteins were activated with similar efficiencies by the individual exchange factors. The H-Ras mutant V103E, which is competent for membrane localization, nucleotide binding, intrinsic and stimulated GTPase activity as well as intrinsic exchange, was defective for activation by all factors tested, suggesting that the integrity of this residue is necessary for catalyzed exchange. However, when other H-Ras mutants were studied, some distinct sensitivities to the exchange factors were observed. GRP-mediated, but not mSos1-mediated, exchange was blocked in additional mutants, suggesting different structural requirements for GRP. Analysis of Ras-mediated gene activation in murine fibroblasts confirmed these results.

Original languageEnglish (US)
Pages (from-to)2091-2100
Number of pages10
JournalOncogene
Volume20
Issue number17
DOIs
StatePublished - Apr 19 2001

Fingerprint

ras Proteins
Alleles
ras Genes
GTP Phosphohydrolases
Transcriptional Activation
Saccharomyces cerevisiae
Nucleotides
Fibroblasts
Membranes
Growth

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Cancer Research
  • Genetics

Cite this

Nielsen, Klaus Hvid ; Gredsted, Lars ; Broach, James ; Willumsen, Berthe Marie. / Sensitivity of wild type and mutant ras alleles to Ras specific exchange factors : Identification of factor specific requirements. In: Oncogene. 2001 ; Vol. 20, No. 17. pp. 2091-2100.
@article{55036b7fe63b4dd2a20f5e57c3c12cb1,
title = "Sensitivity of wild type and mutant ras alleles to Ras specific exchange factors: Identification of factor specific requirements",
abstract = "We have investigated the productive interaction between the four mammalian Ras proteins (H-, N-, KA- and KB-Ras) and their activators, the mammalian exchange factors mSos1, GRF1 and GRP, by using a modified Saccharomyces cerevisiae whose growth is dependent on activation of a mammalian Ras protein by its activator. All four mammalian Ras proteins were activated with similar efficiencies by the individual exchange factors. The H-Ras mutant V103E, which is competent for membrane localization, nucleotide binding, intrinsic and stimulated GTPase activity as well as intrinsic exchange, was defective for activation by all factors tested, suggesting that the integrity of this residue is necessary for catalyzed exchange. However, when other H-Ras mutants were studied, some distinct sensitivities to the exchange factors were observed. GRP-mediated, but not mSos1-mediated, exchange was blocked in additional mutants, suggesting different structural requirements for GRP. Analysis of Ras-mediated gene activation in murine fibroblasts confirmed these results.",
author = "Nielsen, {Klaus Hvid} and Lars Gredsted and James Broach and Willumsen, {Berthe Marie}",
year = "2001",
month = "4",
day = "19",
doi = "10.1038/sj.onc.1204306",
language = "English (US)",
volume = "20",
pages = "2091--2100",
journal = "Oncogene",
issn = "0950-9232",
publisher = "Nature Publishing Group",
number = "17",

}

Sensitivity of wild type and mutant ras alleles to Ras specific exchange factors : Identification of factor specific requirements. / Nielsen, Klaus Hvid; Gredsted, Lars; Broach, James; Willumsen, Berthe Marie.

In: Oncogene, Vol. 20, No. 17, 19.04.2001, p. 2091-2100.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Sensitivity of wild type and mutant ras alleles to Ras specific exchange factors

T2 - Identification of factor specific requirements

AU - Nielsen, Klaus Hvid

AU - Gredsted, Lars

AU - Broach, James

AU - Willumsen, Berthe Marie

PY - 2001/4/19

Y1 - 2001/4/19

N2 - We have investigated the productive interaction between the four mammalian Ras proteins (H-, N-, KA- and KB-Ras) and their activators, the mammalian exchange factors mSos1, GRF1 and GRP, by using a modified Saccharomyces cerevisiae whose growth is dependent on activation of a mammalian Ras protein by its activator. All four mammalian Ras proteins were activated with similar efficiencies by the individual exchange factors. The H-Ras mutant V103E, which is competent for membrane localization, nucleotide binding, intrinsic and stimulated GTPase activity as well as intrinsic exchange, was defective for activation by all factors tested, suggesting that the integrity of this residue is necessary for catalyzed exchange. However, when other H-Ras mutants were studied, some distinct sensitivities to the exchange factors were observed. GRP-mediated, but not mSos1-mediated, exchange was blocked in additional mutants, suggesting different structural requirements for GRP. Analysis of Ras-mediated gene activation in murine fibroblasts confirmed these results.

AB - We have investigated the productive interaction between the four mammalian Ras proteins (H-, N-, KA- and KB-Ras) and their activators, the mammalian exchange factors mSos1, GRF1 and GRP, by using a modified Saccharomyces cerevisiae whose growth is dependent on activation of a mammalian Ras protein by its activator. All four mammalian Ras proteins were activated with similar efficiencies by the individual exchange factors. The H-Ras mutant V103E, which is competent for membrane localization, nucleotide binding, intrinsic and stimulated GTPase activity as well as intrinsic exchange, was defective for activation by all factors tested, suggesting that the integrity of this residue is necessary for catalyzed exchange. However, when other H-Ras mutants were studied, some distinct sensitivities to the exchange factors were observed. GRP-mediated, but not mSos1-mediated, exchange was blocked in additional mutants, suggesting different structural requirements for GRP. Analysis of Ras-mediated gene activation in murine fibroblasts confirmed these results.

UR - http://www.scopus.com/inward/record.url?scp=0035912081&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0035912081&partnerID=8YFLogxK

U2 - 10.1038/sj.onc.1204306

DO - 10.1038/sj.onc.1204306

M3 - Article

C2 - 11360193

AN - SCOPUS:0035912081

VL - 20

SP - 2091

EP - 2100

JO - Oncogene

JF - Oncogene

SN - 0950-9232

IS - 17

ER -