Regular Article
Analysis of Ras-Dependent Signals That Prevent Caspase-3 Activation and Apoptosis Induced by Cytokine Deprivation in Hematopoietic Cells,☆☆

https://doi.org/10.1006/bbrc.1999.1955Get rights and content

Abstract

In hematopoietic cells, Ras has been implicated in signaling pathways that prevent apoptosis triggered by deprivation of cytokines, such as interleukin-3 (IL-3). However, the mechanism whereby Ras suppresses cell death remains incompletely understood. We have investigated the role of Ras in IL-3 signal transduction by using the cytokine-dependent BaF3 cell line. Herein, we show that the activation of the pro-apoptotic protease caspase-3 upon IL-3 removal is suppressed by expression of activated Ras, which eventually prevents cell death. For caspase-3 suppression, the Raf/extracellular signal-regulated kinase (ERK)- or phosphatidylinositol 3-kinase (PI3-K)/Akt-mediated signaling pathway downstream of Ras was required. However, inhibition of both pathways did not block activated Ras-dependent suppression of cell death-associated phenotypes, such as nuclear DNA fragmentation. Thus, a pathway that is independent of both Raf/ERK and PI3-K/Akt pathways may function downstream of Ras, preventing activated caspase-3-initiated apoptotic processes. Conditional activation of c-Raf-1 also suppressed caspase-3 activation and subsequent cell death without affecting Akt activity, providing further evidence for a PI3-K/Akt-independent mechanism.

References (35)

  • K. Terada et al.

    J. Biol. Chem.

    (1995)
  • B.B. Wolf et al.

    J. Biol. Chem.

    (1999)
  • N.L. Harvey et al.

    J. Biol. Chem.

    (1997)
  • T. Ohta et al.

    J. Biol. Chem.

    (1997)
  • X. Liu et al.

    Cell

    (1997)
  • D.D. Newmeyer et al.

    Methods Cell Biol.

    (1991)
  • S. Mitamura et al.

    Biochem. Biophys. Res. Commun.

    (1998)
  • J. Downward

    Curr. Opin. Cell Biol.

    (1998)
  • H. Harada et al.

    Mol. Cell

    (1999)
  • B.L. Craddock et al.

    J. Biol. Chem.

    (1999)
  • A.M. Gardner et al.

    J. Biol. Chem.

    (1996)
  • K. Arai et al.

    Annu. Rev. Biochem.

    (1990)
  • T. Satoh et al.

    Proc. Natl. Acad. Sci. USA

    (1991)
  • V. Duronio et al.

    Proc. Natl. Acad. Sci. USA

    (1992)
  • N. Sato et al.

    EMBO J.

    (1993)
  • T. Kinoshita et al.

    EMBO J.

    (1995)
  • M.S. Marshall

    FASEB J.

    (1995)
  • Cited by (35)

    • Repeated low-dose 17β-estradiol treatment prevents activation of apoptotic signaling both in the synaptosomal and cellular fraction in rat prefrontal cortex following cerebral ischemia

      2015, Neurochemistry International
      Citation Excerpt :

      As ERK and Akt pathways can cross-talk and result in the prevention of apoptosis (Cardone et al., 1998), intrinsic apoptotic pathway may be affected at several different levels. Activation of these pathways inhibits Bax conformational change required for its translocation to the mitochondria (Tsuruta et al., 2002; Yamaguchi and Wang, 2001), Akt may inhibit opening of the mitochondrial permeability transition pore, thereby retaining cytochrome c within the mitochondrial inter membranous space (Hausenloy and Yellon, 2004), while ERK activation has been shown to inhibit apoptosis over inhibition of caspase 3 activation (Terada et al., 2000). In accordance with that, it should not be surprising that increased levels of phospho forms of both ERK (42 kDa pERK and 44 kDa pERK) and Akt (pAkt) were observed in the cytosolic and nuclear fraction of the prefrontal cortex.

    • The protective effect of hispidin against hydrogen peroxide-induced apoptosis in H9c2 cardiomyoblast cells through Akt/GSK-3β and ERK1/2 signaling pathway

      2014, Experimental Cell Research
      Citation Excerpt :

      The activation of either the PI3K/Akt or the ERK1/2 pathway inhibits the conformational change in Bax required for its translocation to the mitochondria, therefore preventing apoptosis [40,41]. Furthermore the up-regulation of ERK1/2 and Akt inactivates the caspase cascade, a proposition which is supported by inhibiting caspase-3 and caspase-9 activation [42,43]. In the present study hispidin increased the phosphorylation of Akt/GSK-3β and ERK1/2 but not p38 MAPK and JNK, which are reduced by H2O2 (Fig. 4).

    • Apurinic/apyrimidinic endonuclease1/redox factor-1 (Ape1/Ref-1) is essential for IL-21-induced signal transduction through ERK1/2 pathway

      2012, Biochemical and Biophysical Research Communications
      Citation Excerpt :

      Further study is needed for this point. Previous reports that showed MEK-ERK activity prevented IL-3 deprivation-induced cell death in BaF3 cells are consistent with our results [35,36]. MEK-ERK pathway plays pivotal roles in the development of various organs and in cell proliferation and survival.

    • Anti-apoptotic pro-survival effect of clotrimazole in a normothermic ischemia reperfusion injury animal model

      2011, Journal of Surgical Research
      Citation Excerpt :

      In an experimental model of reduced size liver transplantation, Liang et al. also found that activation of the ERK1/2 pathway is associated with significantly less apoptosis in the liver [28]. ERK1/2 activation inhibits apoptosis through a conformational change in Bax required for its translocation to the mitochondria [29] and caspase-3 activation [30]. JNK is known to be stimulated in vivo after rat liver transplantation or partial hepatectomy [15, 16].

    • The powerful cardioprotective effects of urocortin and the corticotropin releasing hormone (CRH) family

      2009, Biochemical Pharmacology
      Citation Excerpt :

      This results in it binding to the 14-3-3 protein, thereby preventing it from interacting with mitochondria, and thus conferring resistance against apoptosis [62]. Both MEK1/2 and PI3K are responsible for preventing procaspases 9 and 3 from being cleaved into their active forms [63,64]. Furthermore, chemical inhibitors of the PI3K pathway, such as wortmannin and LY294002, have been reported to block Ucn 1-mediated cardioprotection in both neonatal and adult cardiomyocytes [61].

    View all citing articles on Scopus

    This work was supported in part by CREST of the Japan Science and Technology Corporation. Our laboratory at Tokyo Institute of Technology is supported by Shering-Plough Corporation.

    ☆☆

    Abbreviations used: DTT, dithiothreitol; ERK, extracellular signal-regulated kinase; GM-CSF, granulocyte/macrophage colony-stimulating factor; IL-3, interleukin-3; MEK, mitogen-activated protein kinase/ERK kinase; PI3-K, phosphatidylinositol 3-kinase; PMSF, phenylmethylsulfonyl fluoride

    2

    To whom correspondence should be addressed. Fax: 81-45-924-5822. E-mail: [email protected].

    View full text