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The Journal of Neuroscience, July 1, 2002, 22(13):5393-5402
Gephyrin Interacts with Dynein Light Chains 1 and 2, Components of Motor Protein Complexes
Jens C. Fuhrmann1, Stefan Kins1, Philippe Rostaing2, Oussama El Far1, Joachim Kirsch1, Morgan Sheng3, Antoine Triller2, Heinrich Betz1, and Matthias Kneussel1 1 Max-Planck-Institute for Brain Research, Department of Neurochemistry, D-60528 Frankfurt/Main, Germany, 2 Laboratoire de Biologie Cellulaire de la Synapse, Institut National de la Santé et de la Recherche Médicale U497, Ecole Normale Supérieure, 75005 Paris, France, and 3 Center for Learning and Memory, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139
The clustering of glycine receptors and major subtypes of GABAA receptors at inhibitory synapses is mediated by the tubulin-binding protein gephyrin. In an attempt to identify additional components of inhibitory postsynaptic specializations, we performed a yeast two-hybrid screen using gephyrin as bait. Multiple positive clones encoded either the dynein light chain-1 (Dlc-1), also known as dynein LC8 and protein inhibitor of neuronal nitric oxide synthase, or its homolog Dlc-2. Dlc-1 protein bound efficiently to gephyrin in in vitro binding assays and colocalized with gephyrin during coexpression in HEK293 cells. The binding site for Dlc was mapped to a fragment of 63 amino acids within the central linker domain of gephyrin. In hippocampal neurons, endogenous Dlc protein was enriched at synaptic sites identified by synaptophysin and gephyrin immunostaining. Immunoelectron microscopy in spinal cord sections revealed Dlc immunoreactivity at the edges of postsynaptic differentiations, in close contact with cytoskeletal structures and at the periphery of the Golgi apparatus. Because Dlc-1 and Dlc-2 have been described as stoichiometric components of cytoplasmic dynein and myosin-Va complexes, our results suggest that motor proteins are involved in the subcellular localization of gephyrin.
Key words: gephyrin; cytoplasmic dynein; myosin-Va; Dlc-1; PIN; LC8; Dlc-2; inhibitory synapse; motor protein
Copyright © 2002 Society for Neuroscience 0270-6474/02/22135393-10$05.00/0
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