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Anterograde and retrograde intracellular trafficking of fluorescent cellular prion protein

Naomi S Hachiya1, Kota Watanabe, Makiko Yamada

  • 1Department of Cortical Function Disorders, National Institute of Neuroscience, National Center of Neurology and Psychiatry, and Core Research for Evolutional Science and Technology, Japan Science and Technology Agency, Tokyo 187-8502, Japan.

Biochemical and Biophysical Research Communications
|February 27, 2004
PubMed

Insights

Cellular prion protein (PrPC) moves within cells via microtubule-associated trafficking. Specific N-terminal residues of PrPC are essential for kinesin-driven anterograde and dynein-driven retrograde transport.

Area of Science:

  • Cell Biology
  • Neuroscience
  • Biochemistry

Background:

  • Cellular prion protein (PrPC) is involved in various cellular functions.
  • Intracellular trafficking pathways are crucial for protein localization and function.
  • Microtubule-dependent motor proteins, kinesin and dynein, mediate cargo transport within cells.

Purpose of the Study:

  • To investigate the microtubule-associated intracellular trafficking of the NH2-terminal cellular prion protein (PrPC) fragment.
  • To identify the motor proteins involved in PrPC transport.
  • To map the domains of PrPC responsible for its interaction with motor proteins.

Main Methods:

  • Real-time imaging of fluorescently tagged PrPC (GFP-PrPC) in living cells.
  • Inhibition of motor protein activity using specific inhibitors (AMP-PNP for kinesin, vanadate for dynein).
  • Blocking motor protein function using antibodies (anti-kinesin and anti-dynein).
  • Mapping of interacting domains within the PrPC N-terminus.

Main Results:

  • GFP-PrPC exhibited bidirectional movement: anterograde (140-180 nm/s) and retrograde (1.0-1.2 µm/s).
  • Anterograde movement was inhibited by kinesin inhibitors and antibodies.
  • Retrograde movement was inhibited by dynein inhibitors and antibodies.
  • Specific N-terminal residues (53-91 for kinesin, 23-33 for dynein) were identified as critical for motor protein interaction.

Conclusions:

  • PrPC intracellular trafficking is mediated by both kinesin and dynein motor proteins.
  • Discrete N-terminal amino acid residues of PrPC are indispensable for its anterograde and retrograde movements.
  • These findings provide insights into the mechanism of PrPC transport and its potential role in cellular function.

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