Analysis of the c-src gene product structure, abundance, and protein kinase activity in human neuroblastoma and glioblastoma cells.
We have compared in different human neuroblastoma cell lines and human glioblastoma cells the expression level, structure, and tyrosine-specific protein kinase activity of pp60c-src. Our results show that not all human neuroblastoma cell lines express pp60c-src molecules with amino-terminal structural alterations. In neuroblastoma cells which possess pp60c-src with altered gel migration, the diminished polyacrylamide gel mobility of pp60c-src was found not to be dependent upon amino-terminal phosphorylations since extensive treatment of these molecules with phosphatase did not significantly change their gel migration properties. Similar differences in gel migration were observed when RNA from the various neuroblastoma and glioblastoma cells was translated in vitro using either rabbit reticulocyte or wheat germ lysates. White the level of c-src mRNA in the different cells analyzed was found to be similar, the abundance of pp60c-src in these same cells was found to vary by as much as 12-fold. This suggests that the abundance of pp60c-src in human neuroendocrine tumors is regulated through post-transcriptional and/or post-translational events which may be related to the stage of neuronal differentiation of the cells. Based upon determination of pp60c-src abundance by immunoblot analysis, we demonstrate that pp60c-src molecules derived from human neuroblastoma and glioblastoma cells have very similar in vitro protein kinase activities.
O'Shaughnessy
J
J
Laboratory of Tumor Virus Biology and Medicine Branch, National Cancer Institute, Bethesda, Maryland 20892.
Deseau
V
V
Amini
S
S
Rosen
N
N
Bolen
J B
JB
D000911:Antibodies, Monoclonal; D002460:Cell Line; D004591:Electrophoresis, Polyacrylamide Gel; D005910:Glioma / Q000032:analysis* / Q000235:genetics; D006801:Humans; D015151:Immunoblotting; D009363:Neoplasm Proteins / Q000032:analysis*; D009447:Neuroblastoma / Q000032:analysis* / Q000235:genetics; D015688:Oncogene Protein pp60(v-src); D010766:Phosphorylation; D014176:Protein Biosynthesis; D011505:Protein-Tyrosine Kinases / Q000378:metabolism*; D012191:Retroviridae Proteins / Q000032:analysis*
D000911:Antibodies, Monoclonal; D009363:Neoplasm Proteins; D012191:Retroviridae Proteins; D011505:Protein-Tyrosine Kinases; D015688:Oncogene Protein pp60(v-src)