M-Ras/R-Ras3, a transforming Ras protein regulated by Sos1, GRF1, and p120 Ras GTPase-activating protein, interacts with the putative Ras effector AF6

Lawrence A. Quilliam, Ariel F. Castro, Kelley S. Rogers-Graham, Carol B. Martin, Channing J. Der, Chen Bi

Research output: Contribution to journalArticle

86 Citations (Scopus)

Abstract

M-Ras is a Ras-related protein that shares ~55% identity with K-Ras and TC21. The M-Ras message was widely expressed but was most predominant in ovary and brain. Similarly to Ha-Ras, expression of mutationally activated M- Ras in NIH 3T3 mouse fibroblasts or C2 myoblasts resulted in cellular transformation or inhibition of differentiation, respectively. M-Ras only weakly activated extracellular signal-regulated kinase 2 (ERK2), but it cooperated with Raf, Rac, and Rho to induce transforming foci in NIH 3T3 cells, suggesting that M-Ras signaled via alternate pathways to these effectors. Although the mitogen-activated protein kinase/ERK kinase inhibitor, PD98059, blocked M-Ras-induced transformation, M-Ras was more effective than an activated mitogen-activated protein kinase/ERK kinase mutant at inducing focus formation. These data indicate that multiple pathways must contribute to M-Ras-induced transformation. M-Ras interacted poorly in a yeast two-hybrid assay with multiple Ras effectors, including c- Raf-1, A-Raf, B-Raf, phosphoinositol-3 kinase δ, RalGDS, and Rin1. Although M-Ras coimmunoprecipitated with AF6, a putative regulator of cell junction formation, overexpression of AF6 did not contribute to fibroblast transformation, suggesting the possibility of novel effector proteins. The M- Ras GTP/GDP cycle was sensitive to the Ras GEFs, Sos1, and GRF1 and to p120 Ras GAP. Together, these findings suggest that while M-Ras is regulated by similar upstream stimuli to Ha-Ras, novel targets may be responsible for its effects on cellular transformation and differentiation.

Original languageEnglish (US)
Pages (from-to)23850-23857
Number of pages8
JournalJournal of Biological Chemistry
Volume274
Issue number34
DOIs
StatePublished - Aug 20 1999

Fingerprint

p120 GTPase Activating Protein
ras GTPase-Activating Proteins
ras Proteins
Mitogen-Activated Protein Kinase Kinases
ras-GRF1
ras Guanine Nucleotide Exchange Factors
Phosphotransferases
Fibroblasts
Mitogen-Activated Protein Kinases
NIH 3T3 Cells
Two-Hybrid System Techniques
Intercellular Junctions
Myoblasts
Mitogen-Activated Protein Kinase 1
Guanosine Triphosphate
Ovary
Yeast
Assays
Brain
Proteins

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

M-Ras/R-Ras3, a transforming Ras protein regulated by Sos1, GRF1, and p120 Ras GTPase-activating protein, interacts with the putative Ras effector AF6. / Quilliam, Lawrence A.; Castro, Ariel F.; Rogers-Graham, Kelley S.; Martin, Carol B.; Der, Channing J.; Bi, Chen.

In: Journal of Biological Chemistry, Vol. 274, No. 34, 20.08.1999, p. 23850-23857.

Research output: Contribution to journalArticle

Quilliam, Lawrence A. ; Castro, Ariel F. ; Rogers-Graham, Kelley S. ; Martin, Carol B. ; Der, Channing J. ; Bi, Chen. / M-Ras/R-Ras3, a transforming Ras protein regulated by Sos1, GRF1, and p120 Ras GTPase-activating protein, interacts with the putative Ras effector AF6. In: Journal of Biological Chemistry. 1999 ; Vol. 274, No. 34. pp. 23850-23857.
@article{c2cbe93062db405b87345819af999a78,
title = "M-Ras/R-Ras3, a transforming Ras protein regulated by Sos1, GRF1, and p120 Ras GTPase-activating protein, interacts with the putative Ras effector AF6",
abstract = "M-Ras is a Ras-related protein that shares ~55{\%} identity with K-Ras and TC21. The M-Ras message was widely expressed but was most predominant in ovary and brain. Similarly to Ha-Ras, expression of mutationally activated M- Ras in NIH 3T3 mouse fibroblasts or C2 myoblasts resulted in cellular transformation or inhibition of differentiation, respectively. M-Ras only weakly activated extracellular signal-regulated kinase 2 (ERK2), but it cooperated with Raf, Rac, and Rho to induce transforming foci in NIH 3T3 cells, suggesting that M-Ras signaled via alternate pathways to these effectors. Although the mitogen-activated protein kinase/ERK kinase inhibitor, PD98059, blocked M-Ras-induced transformation, M-Ras was more effective than an activated mitogen-activated protein kinase/ERK kinase mutant at inducing focus formation. These data indicate that multiple pathways must contribute to M-Ras-induced transformation. M-Ras interacted poorly in a yeast two-hybrid assay with multiple Ras effectors, including c- Raf-1, A-Raf, B-Raf, phosphoinositol-3 kinase δ, RalGDS, and Rin1. Although M-Ras coimmunoprecipitated with AF6, a putative regulator of cell junction formation, overexpression of AF6 did not contribute to fibroblast transformation, suggesting the possibility of novel effector proteins. The M- Ras GTP/GDP cycle was sensitive to the Ras GEFs, Sos1, and GRF1 and to p120 Ras GAP. Together, these findings suggest that while M-Ras is regulated by similar upstream stimuli to Ha-Ras, novel targets may be responsible for its effects on cellular transformation and differentiation.",
author = "Quilliam, {Lawrence A.} and Castro, {Ariel F.} and Rogers-Graham, {Kelley S.} and Martin, {Carol B.} and Der, {Channing J.} and Chen Bi",
year = "1999",
month = "8",
day = "20",
doi = "10.1074/jbc.274.34.23850",
language = "English (US)",
volume = "274",
pages = "23850--23857",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "34",

}

TY - JOUR

T1 - M-Ras/R-Ras3, a transforming Ras protein regulated by Sos1, GRF1, and p120 Ras GTPase-activating protein, interacts with the putative Ras effector AF6

AU - Quilliam, Lawrence A.

AU - Castro, Ariel F.

AU - Rogers-Graham, Kelley S.

AU - Martin, Carol B.

AU - Der, Channing J.

AU - Bi, Chen

PY - 1999/8/20

Y1 - 1999/8/20

N2 - M-Ras is a Ras-related protein that shares ~55% identity with K-Ras and TC21. The M-Ras message was widely expressed but was most predominant in ovary and brain. Similarly to Ha-Ras, expression of mutationally activated M- Ras in NIH 3T3 mouse fibroblasts or C2 myoblasts resulted in cellular transformation or inhibition of differentiation, respectively. M-Ras only weakly activated extracellular signal-regulated kinase 2 (ERK2), but it cooperated with Raf, Rac, and Rho to induce transforming foci in NIH 3T3 cells, suggesting that M-Ras signaled via alternate pathways to these effectors. Although the mitogen-activated protein kinase/ERK kinase inhibitor, PD98059, blocked M-Ras-induced transformation, M-Ras was more effective than an activated mitogen-activated protein kinase/ERK kinase mutant at inducing focus formation. These data indicate that multiple pathways must contribute to M-Ras-induced transformation. M-Ras interacted poorly in a yeast two-hybrid assay with multiple Ras effectors, including c- Raf-1, A-Raf, B-Raf, phosphoinositol-3 kinase δ, RalGDS, and Rin1. Although M-Ras coimmunoprecipitated with AF6, a putative regulator of cell junction formation, overexpression of AF6 did not contribute to fibroblast transformation, suggesting the possibility of novel effector proteins. The M- Ras GTP/GDP cycle was sensitive to the Ras GEFs, Sos1, and GRF1 and to p120 Ras GAP. Together, these findings suggest that while M-Ras is regulated by similar upstream stimuli to Ha-Ras, novel targets may be responsible for its effects on cellular transformation and differentiation.

AB - M-Ras is a Ras-related protein that shares ~55% identity with K-Ras and TC21. The M-Ras message was widely expressed but was most predominant in ovary and brain. Similarly to Ha-Ras, expression of mutationally activated M- Ras in NIH 3T3 mouse fibroblasts or C2 myoblasts resulted in cellular transformation or inhibition of differentiation, respectively. M-Ras only weakly activated extracellular signal-regulated kinase 2 (ERK2), but it cooperated with Raf, Rac, and Rho to induce transforming foci in NIH 3T3 cells, suggesting that M-Ras signaled via alternate pathways to these effectors. Although the mitogen-activated protein kinase/ERK kinase inhibitor, PD98059, blocked M-Ras-induced transformation, M-Ras was more effective than an activated mitogen-activated protein kinase/ERK kinase mutant at inducing focus formation. These data indicate that multiple pathways must contribute to M-Ras-induced transformation. M-Ras interacted poorly in a yeast two-hybrid assay with multiple Ras effectors, including c- Raf-1, A-Raf, B-Raf, phosphoinositol-3 kinase δ, RalGDS, and Rin1. Although M-Ras coimmunoprecipitated with AF6, a putative regulator of cell junction formation, overexpression of AF6 did not contribute to fibroblast transformation, suggesting the possibility of novel effector proteins. The M- Ras GTP/GDP cycle was sensitive to the Ras GEFs, Sos1, and GRF1 and to p120 Ras GAP. Together, these findings suggest that while M-Ras is regulated by similar upstream stimuli to Ha-Ras, novel targets may be responsible for its effects on cellular transformation and differentiation.

UR - http://www.scopus.com/inward/record.url?scp=0033588299&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0033588299&partnerID=8YFLogxK

U2 - 10.1074/jbc.274.34.23850

DO - 10.1074/jbc.274.34.23850

M3 - Article

C2 - 10446149

AN - SCOPUS:0033588299

VL - 274

SP - 23850

EP - 23857

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 34

ER -