Mapping the intracytoplasmic regions of the alpha granulocyte-macrophage colony-stimulating factor receptor necessary for cell growth regulation.
The granulocyte-macrophage colony-stimulating factor (GM-CSF) receptor is composed of an alpha subunit which binds GM-CSF and a beta subunit, which together form the high affinity receptor. By transfecting the human alpha subunit into murine Ba/F3 cells, we have been able to investigate the role of the short 54-amino acid intracytoplasmic portion (amino acid 346-400) of this subunit in mediating cell growth. We have shown that the intracytoplasmic amino acids 346-382 are necessary for GM-CSF-mediated cell growth. In contrast, amino acids 382-400 can be removed without effect. The stable transfection of the human beta subunit into the cell lines containing the mutant alpha subunits did not affect the growth characteristics of these cells. The ability of GM-CSF to stimulate cell growth of the Ba/F3 cells alpha subunit transfectants was correlated with the ability of this hormone to translocate protein kinase C to the particulate fraction. In contrast, the ability of GM-CSF addition to increase phosphorylation of the human beta subunit did not correlate with cell growth and required the entire intracytoplasmic domain of the alpha subunit. These results demonstrate an important role for the intracytoplasmic portion of the alpha subunit in mediating both signal transduction and cell cycle commitment stimulated by GM-CSF.
Polotskaya
A
A
Division of Hematology/Oncology, University of Alabama at Birmingham 35294.
Zhao
Y
Y
Lilly
M B
MB
Kraft
A S
AS
D000818:Animals; D001483:Base Sequence; D002455:Cell Division / Q000187:drug effects; D002460:Cell Line; D003001:Cloning, Molecular; D003593:Cytoplasm / Q000378:metabolism; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000378:metabolism* / Q000494:pharmacology*; D006801:Humans; D046911:Macromolecular Substances; D051379:Mice; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009838:Oligodeoxyribonucleotides; D011493:Protein Kinase C / Q000378:metabolism; D016187:Receptors, Granulocyte-Macrophage Colony-Stimulating Factor / Q000096:biosynthesis / Q000737:chemistry / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism / Q000494:pharmacology; D014162:Transfection
D046911:Macromolecular Substances; D009838:Oligodeoxyribonucleotides; D016187:Receptors, Granulocyte-Macrophage Colony-Stimulating Factor; D011994:Recombinant Proteins; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D011493:Protein Kinase C