The unique insert of cellular and viral fms protein tyrosine kinase domains is dispensable for enzymatic and transforming activities.
The receptors for colony stimulating factor-1 (CSF-1), platelet derived growth factor and the c-kit protein tyrosine kinase (PTK) contain within their catalytic domains a stretch of 60-100 residues, largely unrelated in sequence, with no counterpart in other PTKs. Of the 64 amino acids within this kinase insert, 58 were deleted from the mouse CSF-1 receptor by oligonucleotide-directed mutagenesis. The mutant CSF-1 receptor was not markedly affected in its kinase activity, post-translational processing or its ability to induce autocrine transformation of NIH 3T3 mouse fibroblasts. Similarly, retention of kinase and transforming activities were observed following deletion of part or all of the kinase insert from the v-fms oncoprotein. The c- and v-fms kinase inserts were probed using monoclonal and polyclonal antibodies and were found to be highly antigenic. Two monoclonal antibodies raised to the v-fms cytoplasmic domain both recognized epitopes within the insert, and bound enzymatically active v-fms glycoproteins. These results indicate that the fms kinase insert is located on the surface of the protein and folds separately from the rest of the catalytic domain, but is not required for the biological activity of fms PTKs ectopically expressed in mouse fibroblasts. The insert may therefore play a specific function in cells such as monocytes and trophoblasts that normally express the CSF-1 receptor.
Taylor
G R
GR
Division of Molecular and Developmental Biology, Mount Sinai Hospital Research Institute, Toronto, Ontario, Canada.
Reedijk
M
M
Rothwell
V
V
Rohrschneider
L
L
Pawson
T
T
D000595:Amino Acid Sequence; D000818:Animals; D015153:Blotting, Western; D002872:Chromosome Deletion; D004251:DNA Transposable Elements; D005796:Genes; D005814:Genes, Viral; D006023:Glycoproteins / Q000235:genetics*; D051379:Mice; D008969:Molecular Sequence Data; D009154:Mutation; D015345:Oligonucleotide Probes; D016281:Oncogene Protein gp140(v-fms); D011505:Protein-Tyrosine Kinases / Q000235:genetics / Q000378:metabolism*; D015682:Retroviridae Proteins, Oncogenic / Q000235:genetics* / Q000378:metabolism
D004251:DNA Transposable Elements; D006023:Glycoproteins; D015345:Oligonucleotide Probes; D016281:Oncogene Protein gp140(v-fms); D015682:Retroviridae Proteins, Oncogenic; D011505:Protein-Tyrosine Kinases