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Murine c-fms cDNA: cloning, sequence analysis and retroviral expression
V M Rothwell1, L R Rohrschneider
1Cell Biology Department, Fred Hutchinson Cancer Research Center, Seattle, Washington 98104.
Oncogene Research
|September 1, 1987
Summary
Researchers sequenced the murine c-fms gene, revealing a protein with high homology to human and viral forms. Engineered cells expressing this protein specifically bind human CSF-1.
Area of Science:
- Molecular Biology
- Genetics
- Cell Biology
Background:
- The c-fms gene encodes the receptor for colony-stimulating factor 1 (CSF-1).
- Understanding the structure and function of c-fms is crucial for myeloid cell biology.
Purpose of the Study:
- To isolate and sequence the murine c-fms gene.
- To characterize the protein encoded by the murine c-fms gene.
- To investigate the functional properties of the murine c-fms protein in engineered cell lines.
Main Methods:
- cDNA cloning and sequencing of the murine c-fms gene.
- Retroviral vector insertion and G418 selection for gene expression.
- Immunoprecipitation assays to detect c-fms protein.
- Ligand binding assays to assess CSF-1 interaction.
Main Results:
- A 3677-nucleotide cDNA clone for murine c-fms was obtained, coding for a 976-amino acid protein.
- The murine c-fms protein exhibits significant homology to viral (v-fms) and human c-fms proteins, particularly in the cytoplasmic domain.
- Engineered cell lines expressing murine c-fms protein were successfully generated and demonstrated specific binding to human CSF-1.
Conclusions:
- The murine c-fms gene encodes a transmembrane receptor with structural similarities to its human and viral counterparts.
- The cytoplasmic domain shows the highest degree of evolutionary conservation.
- The functional expression of murine c-fms in cell lines validates its role as a CSF-1 receptor and its potential for further study.