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Molecular and Cellular Biology, August 1999, p. 5308-5315, Vol. 19, No. 8
0270-7306/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
B-Raf Inhibits Programmed Cell Death Downstream of Cytochrome
c Release from Mitochondria by Activating the MEK/Erk
Pathway
Peter
Erhardt,
Erin J.
Schremser, and
Geoffrey M.
Cooper*
Department of Biology, Boston University,
Boston, Massachusetts 02215
Received 11 November 1998/Returned for modification 19 January
1999/Accepted 4 May 1999
Growth factor-dependent kinases, such as phosphatidylinositol
3-kinase (PI 3-kinase) and Raf kinases, have been implicated in the
suppression of apoptosis. We have recently established Rat-1
fibroblast cell lines overexpressing B-Raf, leading to
activation of the MEK/Erk mitogen-activated protein kinase pathway.
Overexpression of B-Raf confers resistance to apoptosis
induced by growth factor withdrawal or PI 3-kinase inhibition. This is
accompanied by constitutive activation of Erk without effects on the PI
3-kinase/Akt pathway. The activity of MEK is essential for
cell survival mediated by B-Raf overexpression, since either treatment
with the specific MEK inhibitor PD98059 or expression of
a dominant inhibitory MEK mutant blocks the antiapoptotic
activity of B-Raf. Activation of MEK is not only necessary but also
sufficient for cell survival because overexpression of
constitutively activated MEK, Ras, or Raf-1, like B-Raf, prevents
apoptosis after growth factor deprivation. Overexpression of B-Raf did not interfere with the release of cytochrome c from mitochondria after growth factor
deprivation. However, the addition of cytochrome c to
cytosols of cells overexpressing B-Raf failed to induce caspase
activation. It thus appears that the B-Raf/MEK/Erk pathway confers
protection against apoptosis at the level of cytosolic
caspase activation, downstream of the release of cytochrome
c from mitochondria.
*
Corresponding author. Mailing address: Department of
Biology, Boston University, 5 Cummington St., Boston, MA 02215. Phone: (617) 353-8735. Fax: (617) 353-8484. E-mail:
gmcooper{at}bu.edu.
Molecular and Cellular Biology, August 1999, p. 5308-5315, Vol. 19, No. 8
0270-7306/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
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