Secretion of protease nexin-1 by C6 glioma cells is under the control of a heterotrimeric G protein, Go1.
Heterotrimeric Go proteins have recently been described as regulators of vesicular traffic. The Goalpha gene encodes, by alternative splicing, two Goalpha polypeptides, Go1alpha and Go2alpha. By immunofluorescence and electron microscopy, we detected Go1alpha on the membrane of small intracellular vesicles in C6 glioma cells. After stable transfection of these cells, overexpression of Go1alpha but not Go2alpha was followed by a rise in the secretion of a serine protease inhibitor, protease nexin-1 (PN-1). This secretion was enhanced as a function of the amount of expressed Go1alpha. Metabolic cell labeling indicated that this increase in PN-1 secretion was not the result of an enhancement in PN-1 biosynthesis or a decrease in its uptake, but revealed a potential role of Go1alpha in the regulation of vesicular PN-1 trafficking. Furthermore, activators of Go proteins, mastoparan and a peptide derived from the amino terminus of the growth cone-associated protein GAP43, increased PN-1 secretion in parental and Go1alpha-overexpressing cells. Brefeldin A, an inhibitor of vesicular traffic, inhibited both basal and mastoparan-stimulated PN-1 secretions. These results indicate, that in C6 glioma cells, PN-1 secretion could be regulated by both Go1alpha expression and activation.
Lagriffoul
A
A
CNRS UPR 9023, Centre CNRS-INSERM de Pharmacologie et Endocrinologie, 141 Rue de la Cardonille, F-34094 Montpellier Cedex 5, France. homburg@ccipe.montp.inserm.fr
Charpentier
N
N
Carrette
J
J
Tougard
C
C
Bockaert
J
J
Homburger
V
V
D016564:Amyloid beta-Protein Precursor; D002352:Carrier Proteins / Q000378:metabolism*; D019206:GTP-Binding Protein alpha Subunits, Gi-Go; D019204:GTP-Binding Proteins / Q000378:metabolism*; D005910:Glioma / Q000201:enzymology*; D016244:Guanosine 5'-O-(3-Thiotriphosphate) / Q000494:pharmacology; D006801:Humans; D007700:Kinetics; D015849:Plasminogen Inactivators / Q000378:metabolism*; D058255:Protease Nexins; D011956:Receptors, Cell Surface / Q000378:metabolism; D015842:Serine Proteinase Inhibitors / Q000378:metabolism*; D058258:Serpin E2; D014162:Transfection; D014407:Tumor Cells, Cultured
D016564:Amyloid beta-Protein Precursor; D002352:Carrier Proteins; D015849:Plasminogen Inactivators; D058255:Protease Nexins; D011956:Receptors, Cell Surface; C545486:SERPINE2 protein, human; D015842:Serine Proteinase Inhibitors; D058258:Serpin E2; C052677:protease nexin-protease complex receptor; D016244:Guanosine 5'-O-(3-Thiotriphosphate); D019204:GTP-Binding Proteins; D019206:GTP-Binding Protein alpha Subunits, Gi-Go