Uncoating of clathrin-coated vesicles in presynaptic terminals: roles for Hsc70 and auxilin.
We have examined the roles of Hsc70 and auxilin in the uncoating of clathrin-coated vesicles (CCVs) during neuronal endocytosis. We identified two peptides that inhibit the ability of Hsc70 and auxilin to uncoat CCVs in vitro. When injected into nerve terminals, these peptides inhibited both synaptic transmission and CCV uncoating. Mutation of a conserved HPD motif within the J domain of auxilin prevented binding to Hsc70 in vitro and injecting this mutant protein inhibited CCV uncoating in vivo, demonstrating that the interaction of auxilin with Hsc70 is critical for CCV uncoating. These studies establish that auxilin and Hsc70 participate in synaptic vesicle recycling in neurons and that an interaction between these proteins is required for CCV uncoating.
Morgan
J R
JR
Department of Neurobiology, Duke University Medical Center, Durham, NC 27710, USA.
Prasad
K
K
Jin
S
S
Augustine
G J
GJ
Lafer
E M
EM
D033942:Adaptor Proteins, Vesicular Transport; D000255:Adenosine Triphosphate / Q000494:pharmacology; D000595:Amino Acid Sequence; D000818:Animals; D002352:Carrier Proteins / Q000737:chemistry / Q000494:pharmacology / Q000502:physiology*; D002417:Cattle; D022163:Clathrin-Coated Vesicles / Q000378:metabolism*; D003574:Cytochrome c Group / Q000737:chemistry / Q000494:pharmacology; D004705:Endocytosis; D050883:HSC70 Heat-Shock Proteins; D018840:HSP70 Heat-Shock Proteins / Q000494:pharmacology / Q000502:physiology*; D008565:Membrane Proteins / Q000737:chemistry / Q000494:pharmacology / Q000502:physiology*; D008969:Molecular Sequence Data; D016296:Mutagenesis; D009474:Neurons / Q000648:ultrastructure; D018377:Neurotransmitter Agents / Q000378:metabolism; D010446:Peptide Fragments / Q000494:pharmacology; D017729:Presynaptic Terminals / Q000378:metabolism*; D009435:Synaptic Transmission / Q000187:drug effects
D033942:Adaptor Proteins, Vesicular Transport; D002352:Carrier Proteins; D003574:Cytochrome c Group; D050883:HSC70 Heat-Shock Proteins; D018840:HSP70 Heat-Shock Proteins; D008565:Membrane Proteins; D018377:Neurotransmitter Agents; D010446:Peptide Fragments; D000255:Adenosine Triphosphate