Perverted responses of the human granulocyte-macrophage colony-stimulating factor receptor in mouse cell lines due to cross-species beta-subunit association.
Transfected murine cell lines are commonly used to study the function of many human cytokine or receptor mutants. This study reports the inappropriate activation of the human granulocyte-macrophage colony-stimulating factor (hGM-CSF) receptor by the human GM-CSF antagonist, E21R, when the human receptor is introduced into the murine cell line BaF-B03. E21R-induced proliferation of the BaF-B03 cells is dependent on transfection with both hGM-CSF receptor alpha and beta(c) subunits. Studies on the underlying mechanism revealed constitutive association between human and mouse beta(c) and GM-CSF receptor-alpha, tyrosine phosphorylation of mouse and human beta(c), and association of phosphorylated mouse beta(c) into an activated human GM-CSF receptor complex in response to E21R and GM-CSF. This interspecies receptor cross-talk of receptor signaling subunits may produce misleading results and emphasizes the need to use cell lines devoid of the cognate endogenous receptors for functional analysis of ligand and receptor mutants.
McClure
B
B
Cytokine Receptor Laboratory, Hanson Centre for Cancer Research, Institute of Medical and Veterinary Science, Adelaide, South Australia.
Stomski
F
F
Lopez
A
A
Woodcock
J
J
D000818:Animals; D016477:Artifacts; D002460:Cell Line; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000494:pharmacology*; D006801:Humans; D004915:Leukemia, Erythroblastic, Acute / Q000473:pathology; D007947:Leukemia, Megakaryoblastic, Acute / Q000473:pathology; D008024:Ligands; D046911:Macromolecular Substances; D051379:Mice; D055503:Protein Multimerization; D021122:Protein Subunits; D011505:Protein-Tyrosine Kinases / Q000378:metabolism; D020239:Receptor Cross-Talk; D016187:Receptors, Granulocyte-Macrophage Colony-Stimulating Factor / Q000737:chemistry / Q000187:drug effects* / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000187:drug effects / Q000235:genetics; D011994:Recombinant Proteins; D015398:Signal Transduction; D013045:Species Specificity; D014162:Transfection; D014407:Tumor Cells, Cultured
D008024:Ligands; D046911:Macromolecular Substances; D021122:Protein Subunits; D016187:Receptors, Granulocyte-Macrophage Colony-Stimulating Factor; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; C101659:granulocyte-macrophage colony-stimulating factor, Arg(21)-; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D011505:Protein-Tyrosine Kinases