The role of kinase activity and the kinase insert region in ligand-induced internalization and degradation of the c-fms protein.
Molecular steps in endocytosis and degradation of the c-fms protein were analyzed by following the fate of mutated c-fms molecules after M-CSF binding. A mutant c-fms protein lacking tyrosine kinase activity was rapidly internalized after M-CSF binding but not degraded. Another mutant c-fms molecule that lacked most of the kinase insert region was similarly internalized after M-CSF binding and also not degraded. This indicates that the signal for internalization is separate from that directing degradation of the receptor. It has been shown previously that a c-fms mutant in which the kinase insert domain is deleted retains tyrosine kinase activity but lacks two major sites of autophosphorylation. The degradation step therefore requires both kinase activity and the kinase insert region whereas the internalization step is independent of these factors. The major sites of tyrosine autophosphorylation within the kinase insert region were next mutated to determine whether autophosphorylation in the kinase insert region of c-fms might be the signal that triggers degradation of internalized receptors. These mutant receptors were still rapidly degraded in response to M-CSF. Therefore, ligand-induced degradation of c-fms may require tyrosine phosphorylation of a protein other than the c-fms receptor itself and the kinase insert region may be necessary for recognition of this substrate.
Carlberg
K
K
Fred Hutchinson Cancer Research Center, Seattle, WA 98104.
Tapley
P
P
Haystead
C
C
Rohrschneider
L
L
D000818:Animals; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D007700:Kinetics; D008024:Ligands; D016173:Macrophage Colony-Stimulating Factor / Q000494:pharmacology*; D051379:Mice; D010766:Phosphorylation; D011505:Protein-Tyrosine Kinases / Q000378:metabolism*; D016186:Receptor, Macrophage Colony-Stimulating Factor / Q000235:genetics / Q000378:metabolism*; D013379:Substrate Specificity
D008024:Ligands; D016173:Macrophage Colony-Stimulating Factor; D011505:Protein-Tyrosine Kinases; D016186:Receptor, Macrophage Colony-Stimulating Factor