KIF1Bbeta- And KIF1A-mediated Axonal Transport of Presynaptic Regulator Rab3 Occurs in a GTP-dependent Manner Through DENN/MADD

Nat Cell Biol. 2008 Nov;10(11):1269-79. doi: 10.1038/ncb1785. Epub 2008 Oct 12.

Abstract

Synaptic proteins are synthesized in the cell body and transported down the axon by microtubule-dependent motors. We previously reported that KIF1Bbeta and KIF1A motors are essential for transporting synaptic vesicle precursors; however the mechanisms that regulate transport, as well as cargo recognition and control of cargo loading and unloading remain largely unknown. Here, we show that DENN/MADD (Rab3-GEP) is an essential part of the regulation mechanism through direct interaction with the stalk domain of KIF1Bbeta and KIF1A. We also show that DENN/MADD binds preferentially to GTP-Rab3 and acts as a Rab3 effector. These molecular interactions are fundamental as sequential genetic perturbations revealed that KIF1Bbeta and KIF1A are essential for the transport of DENN/MADD and Rab3, whereas DENN/MADD is essential for the transport of Rab3. GTP-Rab3 was more effectively transported than GDP-Rab3, suggesting that the nucleotide state of Rab3 regulates axonal transport of Rab3-carrying vesicles through preferential interaction with DENN/MADD.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenoviridae / genetics
  • Axonal Transport / physiology*
  • Cell Line
  • Death Domain Receptor Signaling Adaptor Proteins / metabolism*
  • Genetic Vectors
  • Glutathione Transferase / metabolism
  • Guanine Nucleotide Exchange Factors / metabolism*
  • Guanosine Triphosphate / metabolism
  • Immunohistochemistry
  • Kidney / cytology
  • Kinesin / chemistry
  • Kinesin / metabolism*
  • Protein Structure, Tertiary
  • RNA Interference
  • Recombinant Proteins / metabolism
  • Synaptic Vesicles / metabolism
  • Synaptic Vesicles / physiology
  • Transfection
  • rab3 GTP-Binding Proteins / chemistry
  • rab3 GTP-Binding Proteins / genetics
  • rab3 GTP-Binding Proteins / metabolism*

Substances

  • Death Domain Receptor Signaling Adaptor Proteins
  • Guanine Nucleotide Exchange Factors
  • KIF11 protein, human
  • KIF1A protein, human
  • MADD protein, human
  • Recombinant Proteins
  • Guanosine Triphosphate
  • Glutathione Transferase
  • Kinesin
  • rab3 GTP-Binding Proteins