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Ankrd2/ARPP is a novel Akt2 specific substrate and regulates myogenic differentiation upon cellular exposure to

Vittoria Cenni1, Alberto Bavelloni, Francesca Beretti

  • 1IGM-CNR, Unit of Bologna c/o IOR, 40136 Bologna, Italy. vittoria.cenni@cnr.it

Molecular Biology of the Cell
|July 9, 2011
PubMed

Insights

Akt2 specifically phosphorylates Ankrd2 (Ankyrin repeat domain protein 2) at Ser-99, impacting muscle cell differentiation. This Akt-dependent phosphorylation negatively regulates myogenesis under oxidative stress conditions.

Area of Science:

  • Muscle cell biology
  • Cell signaling
  • Proteomics

Background:

  • Akt signaling pathways are vital for muscle cell survival, differentiation, and regeneration.
  • Identifying novel Akt substrates is crucial for understanding muscle physiology.
  • Ankyrin repeat domain protein 2 (Ankrd2) is involved in skeletal muscle stress responses.

Purpose of the Study:

  • To identify novel substrates of Akt in C(2)C(12) murine muscle cells.
  • To investigate the role of Ankrd2 phosphorylation by Akt in muscle cell differentiation.
  • To elucidate the physiological implications of Akt-mediated Ankrd2 phosphorylation.

Main Methods:

  • Proteomic analysis using anti-phospho-Akt substrate antibody.
  • Database searches (Scansite) for predicted Akt substrates.
  • In vitro and in vivo phosphorylation assays (kinase assays, isoform silencing).
  • Oxidative stress induction (H(2)O(2)) and analysis of Ankrd2 phosphorylation.
  • Functional studies using phosphorylation-defective Ankrd2 mutants in C(2)C(12) myoblasts.

Main Results:

  • Ankrd2 was identified as a novel Akt substrate, phosphorylated at Ser-99.
  • Akt2 was confirmed as the specific kinase responsible for Ankrd2 phosphorylation at Ser-99.
  • Oxidative stress induced by H(2)O(2) triggered Ankrd2 phosphorylation by Akt2.
  • Expression of a phosphorylation-defective Ankrd2 mutant accelerated myoblast differentiation.
  • Akt-dependent Ankrd2 phosphorylation negatively regulates myogenesis during stress.

Conclusions:

  • Ankrd2 is a specific substrate of Akt2 in skeletal muscle cells.
  • Akt2-mediated phosphorylation of Ankrd2 at Ser-99 plays a negative regulatory role in myogenesis.
  • This signaling pathway is important for muscle adaptation to oxidative stress conditions.

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