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Rab11-FIP2 interaction with MYO5B regulates movement of Rab11a-containing recycling vesicles

Jenny C Schafer1, Nicholas W Baetz, Lynne A Lapierre

  • 1Section of Surgical Sciences and the Epithelial Biology Center, Vanderbilt University School of Medicine, Nashville, TN, USA.

Traffic (Copenhagen, Denmark)
|December 31, 2013
PubMed

Insights

Specific mutations in Rab11-FIP2 disrupt its interaction with MYO5B, affecting recycling endosome trafficking. This impairs MYO5B tethering, increasing vesicle movement and highlighting the interaction

Area of Science:

  • Cell Biology
  • Molecular Biology
  • Biochemistry

Background:

  • Rab11a, Rab11-FIP2, and MYO5B form a complex regulating recycling endosome trafficking.
  • Understanding the specific molecular interactions within this complex is crucial for elucidating its function.

Purpose of the Study:

  • To define the intermolecular interactions between Rab11-FIP2 and MYO5B necessary for their association.
  • To investigate the functional consequences of disrupting the MYO5B-Rab11-FIP2 interaction on vesicle trafficking.

Main Methods:

  • Random mutagenesis of Rab11-FIP2 to identify critical residues for MYO5B interaction.
  • Yeast two-hybrid assays and co-expression in HeLa cells to assess protein-protein interactions.
  • Confocal microscopy in MDCK and HeLa cells to analyze protein localization and vesicle movement.

Main Results:

  • Specific point mutations (S229P, G233E) in Rab11-FIP2 abolished its interaction with MYO5B in yeast and cell-based assays.
  • Mutations disrupted the colocalization of Rab11-FIP2 with endogenous MYO5B and altered vesicle trafficking dynamics.
  • Perturbation of the MYO5B-Rab11-FIP2 association led to increased vesicle speed and track length, indicating impaired cytoskeletal tethering.

Conclusions:

  • The interaction between MYO5B and Rab11-FIP2 is critical for stabilizing the Rab11a-containing complex.
  • This interaction is essential for regulating the dynamic movements of membrane recycling vesicles by tethering them to the cytoskeleton.
Keywords:
MYO5BRab11Rab11-FIP2membrane recycling

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