A short segment within the cytoplasmic domain of the neural cell adhesion molecule (N-CAM) is essential for N-CAM-induced NF-kappa B activity in astrocytes
Eb. Little et al., A short segment within the cytoplasmic domain of the neural cell adhesion molecule (N-CAM) is essential for N-CAM-induced NF-kappa B activity in astrocytes, P NAS US, 98(5), 2001, pp. 2238-2243
Citations number
47
Categorie Soggetti
Multidisciplinary
Journal title
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
The neural cell adhesion molecule (N-CAM) is expressed on the surface of as
trocytes, where its hemophilic binding leads to the activation of the trans
cription factor NF-kappaB. Transfection of astrocytes with a construct enco
mpassing the transmembrane region and the cytoplasmic domain of N-CAM (desi
gnated Tm-Cyto, amino acids 685-839 in the full-length molecule) inhibited
this activation up to 40%, and inhibited N-CAM-induced translocation of NF-
kappaB to the nucleus. N-CAM also activated NF-kappaB in astrocytes from N-
CAM knockout mice, presumably through binding to a heterophile. This activa
tion, however, was not blocked by Tm-Cyto expression, indicating that the i
nhibitory effect of the Tm-Cyto construct is specific for cell surface N-CA
M. Deletions and point mutations of the cytoplasmic portion of the Tm-Cyto
construct indicated that the region between amino acids 780 and 800 were es
sential for inhibitory activity. This region contains four threonines (788,
793, 794, and 797). Mutation to alanine of T788, T794, or T797, but not T7
93, abolished inhibitory activity, as did mutation of T788 or T797 to aspar
tic acid. A Tm-Cyto construct with T794 mutated to aspartic acid retained i
nhibitory activity but did not itself induce a constitutive NF-kappaB respo
nse. This result suggests that phosphorylation of T794 may be necessary but
is not the triggering event. Overall, these findings define a short segmen
t of the N-CAM cytoplasmic domain that is critical for N-CAM-induced activa
tion of NF-kappaB and may be important in other N-CAM-mediated signaling.