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(Received for publication, December 17, 1996)
From the Department of Biochemistry, School of Medicine, University
of North Carolina, Chapel Hill, North Carolina 27599-7260 and the
Axonal growth cones respond to adhesion molecules
and extracellular matrix components by rapid morphological changes and
growth rate modification. Neurite outgrowth mediated by the neural cell adhesion molecule (NCAM) requires the src family tyrosine
kinase p59fyn in nerve growth cones, but the molecular basis
for this interaction has not been defined. The NCAM140 isoform, which
is found in migrating growth cones, selectively co-immunoprecipitated
with p59fyn from nonionic detergent (Brij 96) extracts of early
postnatal mouse cerebellum and transfected rat B35 neuroblastoma and
COS-7 cells. p59fyn did not associate significantly with
the NCAM180 isoform, which is found at sites of stable neural cell
contacts, or with the glycophosphatidylinositol-linked
NCAM120 isoform. pp60c-src, a tyrosine kinase
that promotes neurite growth on the neuronal cell adhesion
molecule L1, did not interact with any NCAM isoform. Whereas
p59fyn was constitutively associated with NCAM140, the focal
adhesion kinase p125fak, a nonreceptor tyrosine kinase
known to mediate integrin-dependent signaling, became
recruited to the NCAM140-p59fyn complex when cells were reacted
with antibodies against the extracellular region of NCAM. Treatment of
cells with a soluble NCAM fusion protein or with NCAM antibodies caused
a rapid and transient increase in tyrosine phosphorylation of
p125fak and p59fyn. These results
suggest that NCAM140 binding interactions at the cell surface induce
the assembly of a molecular complex of NCAM140, p125fak, and p59fyn and activate the
catalytic function of these tyrosine kinases, initiating a signaling
cascade that may modulate growth cone migration.
Volume 272, Number 13,
Issue of March 28, 1997
pp. 8310-8319
©1997 by The American Society for Biochemistry and Molecular Biology, Inc.
and
Department of Neurobiology, Becton Dickinson Research
Center, Research Triangle Park, North Carolina 27709
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