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Dynamin-related Protein 1 Oligomerization in Solution Impairs Functional Interactions with Membrane-anchored

Ryan W Clinton1, Christopher A Francy1, Rajesh Ramachandran2

  • 1From the Department of Pharmacology, the Center for Mitochondrial Diseases, the Cleveland Center for Membrane and Structural Biology, and.

The Journal of Biological Chemistry
|November 19, 2015
PubMed

Insights

Mitochondrial fission involves dynamin-related protein 1 (Drp1) and mitochondrial fission factor (Mff). Removing Drp1’s variable domain enhances their interaction and GTPase activity, revealing a new mechanism for mitochondrial division.

Area of Science:

  • Cell Biology
  • Molecular Biology
  • Biochemistry

Background:

  • Mitochondrial fission is vital for cellular health, regulated by dynamin-related protein 1 (Drp1).
  • Mitochondrial fission factor (Mff) is an essential partner for Drp1 recruitment to the outer mitochondrial membrane.
  • The precise mechanism of Drp1-Mff interaction and its role in fission remains unclear.

Purpose of the Study:

  • To elucidate the mechanism by which Mff facilitates mitochondrial fission.
  • To investigate the role of Drp1's variable domain and Mff's coiled-coil motif in Drp1-Mff complex formation and activity.
  • To understand how Drp1 and Mff multimeric states regulate their interaction and GTPase activity.

Main Methods:

  • In vitro reconstitution assays using purified Drp1 and Mff proteins.
  • Biochemical assays to measure GTPase activity and protein-protein interactions.
  • Lipid-templated Mff anchoring to mimic physiological membrane environments.

Main Results:

  • Deletion of the Drp1 variable domain (ΔVD Drp1) promotes the formation of a functional Drp1-Mff copolymer with enhanced GTPase activity.
  • Membrane-anchored Mff significantly stimulates GTPase activity for both WT and ΔVD Drp1.
  • Dimeric Drp1 species, not pre-assembled oligomers, are selectively recruited by Mff to initiate fission complex assembly.
  • Mff's coiled-coil motif is not essential for Drp1 binding but enhances Drp1 self-assembly near the membrane.

Conclusions:

  • Drp1-Mff interaction and cooperative GTPase activity are regulated by the multimeric states of both proteins.
  • Dimeric Drp1 is the key species recruited by Mff to initiate mitochondrial fission.
  • This study provides a mechanistic framework for Drp1-Mff collaboration in mitochondrial division.
Keywords:
GTPasedynamindynamin-related protein 1electron microscopy (EM)membrane remodelingmitochondriamitochondrial fissionmitochondrial fission factorprotein-protein interaction

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