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Akt Regulates a Rab11-Effector Switch Required for Ciliogenesis.

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Summary

Serum growth factors inhibit cilia formation by activating Akt signaling, which blocks key protein interactions. Disrupting Akt signaling promotes ciliogenesis by enabling essential protein complexes to traffic to the mother centriole.

Keywords:
AktFIP3LPAMCRab11 effector switchRabin8WDR44ciliogenesislysophosphatidic acidmother centriolephosphorylationpreciliary trafficking

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Area of Science:

  • Cell Biology
  • Molecular Biology
  • Signaling Pathways

Background:

  • Serum starvation promotes cilia growth, but the specific serum factors involved remain unclear.
  • Previous work identified Rab11-dependent trafficking of Rabin8 (a Rab8 GEF) to the mother centriole as crucial for ciliogenesis.
  • The precise molecular mechanisms by which serum signals regulate this process are not fully understood.

Purpose of the Study:

  • To elucidate the role of serum lysophosphatidic acid (LPA) and its receptor (LPAR1) in regulating ciliogenesis.
  • To investigate the downstream signaling pathways, including PI3K/Akt, involved in serum-mediated inhibition of cilia growth.
  • To understand how Akt signaling impacts the interaction of key proteins like Rab11a, Rabin8, and WDR44 during ciliogenesis.

Main Methods:

  • Cell culture and serum starvation protocols.
  • Analysis of protein-protein interactions using co-immunoprecipitation.
  • Western blotting to assess protein phosphorylation and activation (e.g., Akt).
  • Depletion studies using siRNA to investigate the function of WDR44.
  • Microscopy to observe cilia growth and protein localization.

Main Results:

  • Serum LPA, acting through LPAR1, inhibits ciliogenesis by preventing the Rab11a-Rabin8 interaction.
  • LPA/LPAR1 signaling activates PI3K/Akt, which independently of cell cycle effects, inhibits ciliogenesis.
  • Akt stabilizes the binding of Rab11a to WDR44, and a phosphomimetic WDR44 mutant blocks ciliogenesis.
  • WDR44 depletion enhances Rabin8 preciliary trafficking and promotes ciliogenesis.
  • Akt signaling disruption shifts the balance from the Rab11-WDR44 complex to the ciliogenic Rab11-FIP3-Rabin8 complex.

Conclusions:

  • Serum mitogen signaling, via LPA/LPAR1 and PI3K/Akt, actively inhibits ciliogenesis initiation.
  • Akt signaling regulates ciliogenesis by controlling the preciliary trafficking of Rabin8 through modulation of Rab11a-WDR44 interactions.
  • This study reveals a novel mechanism linking serum growth factor signaling to the regulation of Rab8-dependent cilia formation.