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The Journal of Neuroscience, August 1, 1999, 19(15):6519-6527
Cloning and Characterization of Neuropilin-1-Interacting
Protein: A PSD-95/Dlg/ZO-1 Domain-Containing Protein That
Interacts with the Cytoplasmic Domain of Neuropilin-1
Huaibin
Cai1, 2 and
Randall R.
Reed1, 2, 3
1 Howard Hughes Medical Institutes and Departments of
2 Neuroscience and 3 Molecular Biology and
Genetics, The Johns Hopkins University School of Medicine, Baltimore,
Maryland 21205
Neuropilin-1 (Npn-1), a receptor for semaphorin III, mediates the
guidance of growth cones on extending neurites. The molecular mechanism
of Npn-1 signaling remains unclear. We have used a yeast two-hybrid
system to isolate a protein that interacts with the cytoplasmic domain
of Npn-1. This Npn-1-interacting protein (NIP) contains a central
PSD-95/Dlg/ZO-1 (PDZ) domain and a C-terminal acyl carrier
protein domain. The physiological interaction of Npn-1 and NIP is
supported by co-immunoprecipitation of these two proteins in extracts
from a heterologous expression system and from a native tissue. The
C-terminal three amino acids of Npn-1 (S-E-A-COOH), which is conserved
from Xenopus to human, is responsible for interaction
with the PDZ domain-containing C-terminal two-thirds of NIP. NIP as
well as Npn-1 are broadly expressed in mice as assayed by Northern and
Western analysis. Immunohistochemistry and in situ
hybridization experiments revealed that NIP expression overlaps with
that of Npn-1. NIP has been independently cloned as
RGS-GAIP-interacting protein (GIPC), where it was identified by
virtue of its interaction with the C terminus of RGS-GAIP and
suggested to participate in clathrin-coated vesicular trafficking. We
suggest that NIP and GIPC may participate in regulation of
Npn-1-mediated signaling as a molecular adapter that couples Npn-1 to
membrane trafficking machinery in the dynamic axon growth cone.
Key words:
neuropilin-1; PDZ domain; axon guidance; yeast
two-hybrid; neuron development; adapter protein; signal
transduction
Copyright © 1999 Society for Neuroscience 0270-6474/99/19156519-09$05.00/0
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