Neuroblastoma differentiation involves the expression of two isoforms of the alpha-subunit of Go.
The regulation of GTP-binding proteins (G proteins) was examined during the course of differentiation of neuroblastoma N1E-115 cells. N1E-115 cell membranes possess three Bordetella pertussis toxin (PTX) substrates assigned to alpha-subunits (G alpha) of Go (a G protein of unknown function) and "Gi (a G protein inhibitory to adenylate cyclase)-like" proteins and one substrate of Vibrio cholerae toxin corresponding to an alpha-subunit of Gs (a G protein stimulatory to adenylate cyclase). In undifferentiated cells, only one form of Go alpha was found, having a pI of 5.8 Go alpha content increased by approximately twofold from the undifferentiated state to 96 h of cell differentiation. This is mainly due to the appearance of another Go alpha form having a pI of 5.55. Both Go alpha isoforms have similar sizes on sodium dodecyl sulfate-polyacrylamide gels, are recognized by polyclonal antibodies to bovine brain Go alpha, are ADP-ribosylated by PTX, and are covalently myristylated in whole N1E-115 cells. In addition, immunofluorescent staining of N1E-115 cells with Go alpha antibodies revealed that association of Go alpha with the plasma membrane appears to coincide with the expression of the most acidic isoform and morphological cell differentiation. In contrast, the levels of both Gi alpha and Gs alpha did not significantly change, whereas that of the common beta-subunit increased by approximately 30% over the same period. These results demonstrate specific regulation of the expression of Go alpha during neuronal differentiation.
Brabet
P
P
Centre CNRS-INSERM de Pharmacologie et Endocrinologie, Montpellier, France.
Pantaloni
C
C
Rodriguez
M
M
Martinez
J
J
Bockaert
J
J
Homburger
V
V
D037361:Adenylate Cyclase Toxin; D002454:Cell Differentiation; D002540:Cerebral Cortex / Q000166:cytology / Q000378:metabolism; D019204:GTP-Binding Proteins / Q000145:classification / Q000378:metabolism*; D015151:Immunoblotting; D007150:Immunohistochemistry; D007525:Isoelectric Focusing; D007536:Isomerism; D019814:Myristic Acid; D009227:Myristic Acids / Q000378:metabolism; D009447:Neuroblastoma / Q000378:metabolism / Q000473:pathology*; D037342:Pertussis Toxin; D014018:Tissue Distribution; D014407:Tumor Cells, Cultured; D010566:Virulence Factors, Bordetella / Q000378:metabolism
D037361:Adenylate Cyclase Toxin; D009227:Myristic Acids; D010566:Virulence Factors, Bordetella; D019814:Myristic Acid; D037342:Pertussis Toxin; D019204:GTP-Binding Proteins