Protein kinase A regulates caspase-9 activation by Apaf-1 downstream of cytochrome c

Morag C. Martin, Lindsey A. Allan, Michelle Lickrish, Catherine Sampson, Nick Morrice, Paul R. Clarke (Lead / Corresponding author)

Research output: Contribution to journalArticle

79 Citations (Scopus)

Abstract

The cyclic AMP signal transduction pathway modulates apoptosis in diverse cell types, although the mechanism is poorly understood. A critical component of the intrinsic apoptotic pathway is caspase-9, which is activated by Apaf-1 in the apoptosome, a large complex assembled in response to release of cytochrome c from mitochondria. Caspase-9 cleaves and activates effector caspases, predominantly caspase-3, resulting in the demise of the cell. Here we identified a distinct mechanism by which cyclic AMP regulates this apoptotic pathway through activation of protein kinase A. We show that protein kinase A inhibits activation of caspase-9 and caspase-3 downstream of cytochrome c in Xenopus egg extracts and in a human cell-free system. Protein kinase A directly phosphorylates human caspase-9 at serines 99, 183, and 195. However, mutational analysis demonstrated that phosphorylation at these sites is not required for the inhibitory effect of protein kinase A on caspase-9 activation. Importantly, protein kinase A inhibits cytochrome c-dependent recruitment of pro-caspase-9 to Apaf-1 but not activation of caspase-9 by a constitutively activated form of Apaf-1. These data indicate that extracellular signals that elevate cyclic AMP and activate protein kinase A may suppress apoptosis by inhibiting apoptosome formation downstream of cytochrome c release from mitochondria.

Original languageEnglish
Pages (from-to)15449-15455
Number of pages7
JournalJournal of Biological Chemistry
Volume280
Issue number15
DOIs
Publication statusPublished - 15 Apr 2005

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Cytochromes c1
Caspase 9
Cyclic AMP-Dependent Protein Kinases
Cytochromes c
Chemical activation
Apoptosomes
Cyclic AMP
Mitochondria
Caspase 3
Effector Caspases
Apoptosis
Signal transduction
Phosphorylation
Cell-Free System
Xenopus
Serine
Ovum
Signal Transduction
Cells

Cite this

Martin, Morag C. ; Allan, Lindsey A. ; Lickrish, Michelle ; Sampson, Catherine ; Morrice, Nick ; Clarke, Paul R. / Protein kinase A regulates caspase-9 activation by Apaf-1 downstream of cytochrome c. In: Journal of Biological Chemistry. 2005 ; Vol. 280, No. 15. pp. 15449-15455.
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Protein kinase A regulates caspase-9 activation by Apaf-1 downstream of cytochrome c. / Martin, Morag C.; Allan, Lindsey A.; Lickrish, Michelle; Sampson, Catherine; Morrice, Nick; Clarke, Paul R. (Lead / Corresponding author).

In: Journal of Biological Chemistry, Vol. 280, No. 15, 15.04.2005, p. 15449-15455.

Research output: Contribution to journalArticle

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