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Membrane insertion of the pleckstrin homology domain variable loop 1 is critical for dynamin-catalyzed vesicle

Rajesh Ramachandran1, Thomas J Pucadyil, Ya-Wen Liu

  • 1Department of Cell Biology, The Scripps Research Institute, La Jolla, CA 92037, USA. rxr275@case.edu

Molecular Biology of the Cell
|September 25, 2009
PubMed

Insights

The hydrophobic insertion of dynamin's PH domain is crucial for membrane fission during clathrin-mediated endocytosis. This process, essential for vesicle release, requires both dynamin self-assembly and PH domain membrane interaction.

Area of Science:

  • Cell Biology
  • Molecular Biology
  • Membrane Trafficking

Background:

  • Dynamin (a GTPase) mediates membrane scission during clathrin-coated pit formation.
  • The precise mechanisms of dynamin-driven membrane fission are not fully understood.

Purpose of the Study:

  • To investigate the role of the pleckstrin homology (PH) domain's variable loop 1 (VL1) in dynamin-mediated membrane fission.
  • To determine the importance of PH domain membrane insertion for generating membrane curvature and facilitating vesicle release.

Main Methods:

  • Site-directed mutagenesis to alter the hydrophobic nature of dynamin-1's PH domain VL1.
  • In vitro assays using precurved membrane templates.
  • In vivo studies of clathrin-mediated endocytosis.

Main Results:

  • Stable insertion of the PH domain VL1 into the lipid bilayer is critical for generating high membrane curvature.
  • Mutants defective in curvature generation can be rescued on precurved membranes in vitro.
  • These mutants remain defective in scission of clathrin-coated pits in vivo.

Conclusions:

  • PH domain membrane insertion, alongside dynamin self-assembly, is essential for membrane fission and vesicle release.
  • This mechanism is vital for efficient clathrin-mediated endocytosis in vivo.

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