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

Rajesh Ramachandran, Thomas J. Pucadyil, Ya Wen Liu, Sharmistha Acharya, Marilyn Leonard, Vasyl Lukiyanchuk, Sandra L. Schmid

Research output: Contribution to journalArticle

72 Citations (Scopus)

Abstract

The GTPase dynamin catalyzes the scission of deeply invaginated clathrin-coated pits at the plasma membrane, but the mechanisms governing dynamin-mediated membrane fission remain poorly understood. Through mutagenesis, we have altered the hydrophobic nature of the membrane-inserting variable loop 1 (VL1) of the pleckstrin homology (PH) domain of dynamin-1 and demonstrate that its stable insertion into the lipid bilayer is critical for high membrane curvature generation and subsequent membrane fission. Dynamin PH domain mutants defective in curvature generation regain function when assayed on precurved membrane templates in vitro, but they remain defective in the scission of clathrin-coated pits in vivo. These results demonstrate that, in concert with dynamin self-assembly, PH domain membrane insertion is essential for fission and vesicle release in vitro and for clathrin-mediated endocytosis in vivo.

Original languageEnglish (US)
Pages (from-to)4630-4639
Number of pages10
JournalMolecular Biology of the Cell
Volume20
Issue number22
DOIs
StatePublished - 2009

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Dynamins
Clathrin
Membranes
Dynamin I
GTP Phosphohydrolases
Lipid Bilayers
Endocytosis
Mutagenesis
Pleckstrin Homology Domains
Cell Membrane

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

Cite this

Membrane insertion of the pleckstrin homology domain variable loop 1 is critical for dynamin-catalyzed vesicle scission. / Ramachandran, Rajesh; Pucadyil, Thomas J.; Liu, Ya Wen; Acharya, Sharmistha; Leonard, Marilyn; Lukiyanchuk, Vasyl; Schmid, Sandra L.

In: Molecular Biology of the Cell, Vol. 20, No. 22, 2009, p. 4630-4639.

Research output: Contribution to journalArticle

Ramachandran, Rajesh ; Pucadyil, Thomas J. ; Liu, Ya Wen ; Acharya, Sharmistha ; Leonard, Marilyn ; Lukiyanchuk, Vasyl ; Schmid, Sandra L. / Membrane insertion of the pleckstrin homology domain variable loop 1 is critical for dynamin-catalyzed vesicle scission. In: Molecular Biology of the Cell. 2009 ; Vol. 20, No. 22. pp. 4630-4639.
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