α-thrombin induces rapid and sustained Akt phosphorylation by β-arrestin1-dependent and -independent mechanisms, and only the sustained Akt phosphorylation is essential for G1 phase progression

Reema Goel, Polly J. Phillips-Mason, Daniel M. Raben, Joseph J. Baldassare

Research output: Contribution to journalArticle

78 Citations (Scopus)

Abstract

In Chinese hamster embryonic fibroblasts (IIC9 cells) α-thrombin activates the MAPK(ERK) and phosphatidyl-inositol 3-OH-kinase (PI 3-kinase)/Akt pathways, and both are essential for progression through the G1, phase of the cell cycle. We investigated in IIC9 cells, the role of β-arrestin1 in α-thrombin signaling to these pathways. α-Thrombin stimulates rapid and sustained PI 3-kinase and Akt activities. Expression of a dominant negative 13-arrestin1 (β-arrestin1(V53D)) inhibits rapid but not sustained PI 3-kinase and Akt activities. Surprisingly, expression of β-arrestin1(V53D) does not block activation of the MAPK(ERK) pathway. PI 3-kinase and Akt activities are also inhibited by expression of a β-arrestin1 mutant, which impairs binding to c-Src (β-arrestin1(P91G-P121E)), indicating the involvement of c-Src in the rapid stimulation of the PI 3-kinase/Akt pathway. Consistent with these results, PP1, a selective inhibitor of c-Src family kinases, prevents α-thrombin-stimulated Akt phosphorylation. Expression of β-arrestin1(V53D) does not prevent G1 progression, as its expression has no effect on [3H]thymidine incorporation into DNA. In agreement with the ineffectiveness of β-arrestin1(V53D) to block G1, progression, cyclin D1 protein amounts and CDK4-cyclin D1 activity is unaffected by expression of β-arrestin1(V53D). Thus in IIC9 cells, α-thrombin activates rapid β-arrestin1-dependent and sustained β-arrestin1-independent Akt activity, suggesting that two mechanisms are involved. Furthermore, although blocking the β-arrestin1-independent PI 3-kinase/Akt pathway prevents G1, progression, inhibition of the β-arrestin1-dependent pathway does not, indicating different roles for the rapid and sustained activities.

Original languageEnglish (US)
Pages (from-to)18640-18648
Number of pages9
JournalJournal of Biological Chemistry
Volume277
Issue number21
DOIs
StatePublished - May 24 2002

Fingerprint

Phosphorylation
G1 Phase
Phosphatidylinositols
Thrombin
Phosphotransferases
Cyclin D1
src-Family Kinases
MAP Kinase Signaling System
Fibroblasts
Cricetulus
Thymidine
hydroxide ion
Cell Cycle
Chemical activation
Cells
DNA
Proteins

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

@article{1a0dde0474894c19937180351d7890f1,
title = "α-thrombin induces rapid and sustained Akt phosphorylation by β-arrestin1-dependent and -independent mechanisms, and only the sustained Akt phosphorylation is essential for G1 phase progression",
abstract = "In Chinese hamster embryonic fibroblasts (IIC9 cells) α-thrombin activates the MAPK(ERK) and phosphatidyl-inositol 3-OH-kinase (PI 3-kinase)/Akt pathways, and both are essential for progression through the G1, phase of the cell cycle. We investigated in IIC9 cells, the role of β-arrestin1 in α-thrombin signaling to these pathways. α-Thrombin stimulates rapid and sustained PI 3-kinase and Akt activities. Expression of a dominant negative 13-arrestin1 (β-arrestin1(V53D)) inhibits rapid but not sustained PI 3-kinase and Akt activities. Surprisingly, expression of β-arrestin1(V53D) does not block activation of the MAPK(ERK) pathway. PI 3-kinase and Akt activities are also inhibited by expression of a β-arrestin1 mutant, which impairs binding to c-Src (β-arrestin1(P91G-P121E)), indicating the involvement of c-Src in the rapid stimulation of the PI 3-kinase/Akt pathway. Consistent with these results, PP1, a selective inhibitor of c-Src family kinases, prevents α-thrombin-stimulated Akt phosphorylation. Expression of β-arrestin1(V53D) does not prevent G1 progression, as its expression has no effect on [3H]thymidine incorporation into DNA. In agreement with the ineffectiveness of β-arrestin1(V53D) to block G1, progression, cyclin D1 protein amounts and CDK4-cyclin D1 activity is unaffected by expression of β-arrestin1(V53D). Thus in IIC9 cells, α-thrombin activates rapid β-arrestin1-dependent and sustained β-arrestin1-independent Akt activity, suggesting that two mechanisms are involved. Furthermore, although blocking the β-arrestin1-independent PI 3-kinase/Akt pathway prevents G1, progression, inhibition of the β-arrestin1-dependent pathway does not, indicating different roles for the rapid and sustained activities.",
author = "Reema Goel and Phillips-Mason, {Polly J.} and Raben, {Daniel M.} and Baldassare, {Joseph J.}",
year = "2002",
month = "5",
day = "24",
doi = "10.1074/jbc.M108995200",
language = "English (US)",
volume = "277",
pages = "18640--18648",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "21",

}

α-thrombin induces rapid and sustained Akt phosphorylation by β-arrestin1-dependent and -independent mechanisms, and only the sustained Akt phosphorylation is essential for G1 phase progression. / Goel, Reema; Phillips-Mason, Polly J.; Raben, Daniel M.; Baldassare, Joseph J.

In: Journal of Biological Chemistry, Vol. 277, No. 21, 24.05.2002, p. 18640-18648.

Research output: Contribution to journalArticle

TY - JOUR

T1 - α-thrombin induces rapid and sustained Akt phosphorylation by β-arrestin1-dependent and -independent mechanisms, and only the sustained Akt phosphorylation is essential for G1 phase progression

AU - Goel, Reema

AU - Phillips-Mason, Polly J.

AU - Raben, Daniel M.

AU - Baldassare, Joseph J.

PY - 2002/5/24

Y1 - 2002/5/24

N2 - In Chinese hamster embryonic fibroblasts (IIC9 cells) α-thrombin activates the MAPK(ERK) and phosphatidyl-inositol 3-OH-kinase (PI 3-kinase)/Akt pathways, and both are essential for progression through the G1, phase of the cell cycle. We investigated in IIC9 cells, the role of β-arrestin1 in α-thrombin signaling to these pathways. α-Thrombin stimulates rapid and sustained PI 3-kinase and Akt activities. Expression of a dominant negative 13-arrestin1 (β-arrestin1(V53D)) inhibits rapid but not sustained PI 3-kinase and Akt activities. Surprisingly, expression of β-arrestin1(V53D) does not block activation of the MAPK(ERK) pathway. PI 3-kinase and Akt activities are also inhibited by expression of a β-arrestin1 mutant, which impairs binding to c-Src (β-arrestin1(P91G-P121E)), indicating the involvement of c-Src in the rapid stimulation of the PI 3-kinase/Akt pathway. Consistent with these results, PP1, a selective inhibitor of c-Src family kinases, prevents α-thrombin-stimulated Akt phosphorylation. Expression of β-arrestin1(V53D) does not prevent G1 progression, as its expression has no effect on [3H]thymidine incorporation into DNA. In agreement with the ineffectiveness of β-arrestin1(V53D) to block G1, progression, cyclin D1 protein amounts and CDK4-cyclin D1 activity is unaffected by expression of β-arrestin1(V53D). Thus in IIC9 cells, α-thrombin activates rapid β-arrestin1-dependent and sustained β-arrestin1-independent Akt activity, suggesting that two mechanisms are involved. Furthermore, although blocking the β-arrestin1-independent PI 3-kinase/Akt pathway prevents G1, progression, inhibition of the β-arrestin1-dependent pathway does not, indicating different roles for the rapid and sustained activities.

AB - In Chinese hamster embryonic fibroblasts (IIC9 cells) α-thrombin activates the MAPK(ERK) and phosphatidyl-inositol 3-OH-kinase (PI 3-kinase)/Akt pathways, and both are essential for progression through the G1, phase of the cell cycle. We investigated in IIC9 cells, the role of β-arrestin1 in α-thrombin signaling to these pathways. α-Thrombin stimulates rapid and sustained PI 3-kinase and Akt activities. Expression of a dominant negative 13-arrestin1 (β-arrestin1(V53D)) inhibits rapid but not sustained PI 3-kinase and Akt activities. Surprisingly, expression of β-arrestin1(V53D) does not block activation of the MAPK(ERK) pathway. PI 3-kinase and Akt activities are also inhibited by expression of a β-arrestin1 mutant, which impairs binding to c-Src (β-arrestin1(P91G-P121E)), indicating the involvement of c-Src in the rapid stimulation of the PI 3-kinase/Akt pathway. Consistent with these results, PP1, a selective inhibitor of c-Src family kinases, prevents α-thrombin-stimulated Akt phosphorylation. Expression of β-arrestin1(V53D) does not prevent G1 progression, as its expression has no effect on [3H]thymidine incorporation into DNA. In agreement with the ineffectiveness of β-arrestin1(V53D) to block G1, progression, cyclin D1 protein amounts and CDK4-cyclin D1 activity is unaffected by expression of β-arrestin1(V53D). Thus in IIC9 cells, α-thrombin activates rapid β-arrestin1-dependent and sustained β-arrestin1-independent Akt activity, suggesting that two mechanisms are involved. Furthermore, although blocking the β-arrestin1-independent PI 3-kinase/Akt pathway prevents G1, progression, inhibition of the β-arrestin1-dependent pathway does not, indicating different roles for the rapid and sustained activities.

UR - http://www.scopus.com/inward/record.url?scp=0037166254&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0037166254&partnerID=8YFLogxK

U2 - 10.1074/jbc.M108995200

DO - 10.1074/jbc.M108995200

M3 - Article

C2 - 11901145

AN - SCOPUS:0037166254

VL - 277

SP - 18640

EP - 18648

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 21

ER -