Regulation of intermediary metabolism by the PKCdelta signalosome in mitochondria. Academic Article uri icon

Overview

MeSH

  • Animals
  • Cells, Cultured
  • Cytochromes c
  • Immunoblotting
  • Mice
  • Mice, Inbred C57BL
  • Mice, Knockout
  • Oxidative Phosphorylation
  • Protein-Serine-Threonine Kinases
  • Pyruvate Decarboxylase
  • Shc Signaling Adaptor Proteins
  • Signal Transduction
  • Src Homology 2 Domain-Containing, Transforming Protein 1
  • Vitamin A

MeSH Major

  • Mitochondria
  • Multiprotein Complexes
  • Protein Kinase C-delta

abstract

  • PKCδ has emerged as a novel regulatory molecule of oxidative phosphorylation by targeting the pyruvate dehydrogenase complex (PDHC). We showed that activation of PKCδ leads to the dephosphorylation of pyruvate dehydrogenase kinase 2 (PDK2), thereby decreasing PDK2 activity and increasing PDH activity, accelerating oxygen consumption, and augmenting ATP synthesis. However, the molecular components that mediate PKCδ signaling in mitochondria have remained elusive so far. Here, we identify for the first time a functional complex, which includes cytochrome c as the upstream driver of PKCδ, and uses the adapter protein p66Shc as a platform with vitamin A (retinol) as a fourth partner. All four components are necessary for the activation of the PKCδ signal chain. Genetic ablation of any one of the three proteins, or retinol depletion, silences signaling. Furthermore, mutations that disrupt the interaction of cytochrome c with p66Shc, of p66Shc with PKCδ, or the deletion of the retinol-binding pocket on PKCδ, attenuate signaling. In cytochrome c-deficient cells, reintroduction of cytochrome c Fe(3+) protein restores PKCδ signaling. Taken together, these results indicate that oxidation of PKCδ is key to the activation of the pathway. The PKCδ/p66Shc/cytochrome c signalosome might have evolved to effect site-directed oxidation of zinc-finger structures of PKCδ, which harbor the activation centers and the vitamin A binding sites. Our findings define the molecular mechanisms underlying the signaling function of PKCδ in mitochondria.

publication date

  • December 2010

has subject area

  • Animals
  • Cells, Cultured
  • Cytochromes c
  • Immunoblotting
  • Mice
  • Mice, Inbred C57BL
  • Mice, Knockout
  • Mitochondria
  • Multiprotein Complexes
  • Oxidative Phosphorylation
  • Protein Kinase C-delta
  • Protein-Serine-Threonine Kinases
  • Pyruvate Decarboxylase
  • Shc Signaling Adaptor Proteins
  • Signal Transduction
  • Src Homology 2 Domain-Containing, Transforming Protein 1
  • Vitamin A

Research

keywords

  • Journal Article

Identity

Language

  • eng

PubMed Central ID

  • PMC2992363

Digital Object Identifier (DOI)

  • 10.1096/fj.10-166934

PubMed ID

  • 20798245

Additional Document Info

start page

  • 5033

end page

  • 5042

volume

  • 24

number

  • 12