The c-fps/fes gene encodes a cytoplasmic tyrosine kinase that is specifically expressed in myeloid cells, in particular in macrophages. The long-term objective of this study is to understand the biological role and mechanism of action of this gene, and the molecular basis of its oncogenic potential. We will investigate the possible role of c-fps/fes in signal transduction by determining if its product undergoes any biochemical changes (i.e. tyrosine phosphorylation and activation of enzymatic activity) during myeloid cell differentiation and macrophage activation, and by examining the biological and biochemical effects of reintroducing biologically active fps/fes genes into these cells. The biochemical parameters of c-fps/fes action will be dissected using temperature sensitive and other mutants of c-fps/fes, which will be introduced into myeloid cells capable of undergoing differentiation and maturation by retroviral mediated gene transfer. These experiments should clarify the biological functions of c-fps/fes and provide the means to identify proliferation-, differentiation-, and maturation-specific substrates. We will also generate antibodies against two previously identified substrates of c-fps/fes, which will be used for their characterization and molecular cloning. We will identify cellular proteins that interact with c-fps/fes kinase using purified c-fps/fes protein expressed in baculovirus, and functional domains expressed in bacteria, as affinity reagents. These proteins will also be used to generate specific monoclonal and polyclonal antibodies, which will be used in conjunction with fps/fes mutants to study functional interactions of c-fps/fes kinase with substrates and other cellular proteins. The c-fps/fes product has unique biological and biochemical properties. The elucidation of its mechanism of action will shed light on the role of tyrosine phosphorylation in myeloid cell differentiation and macrophage activation. As macrophage activation plays a central role in the immune response to infections, and as new experimental therapies rely on the use of myeloid CSFs that utilize tyrosine phosphorylation pathways to achieve differentiation of leukemic cells, the mechanisms uncovered in these studies have broad potential clinical implications.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
First Independent Research Support & Transition (FIRST) Awards (R29)
Project #
5R29CA055293-04
Application #
2096505
Study Section
Cellular Biology and Physiology Subcommittee 1 (CBY)
Project Start
1991-07-01
Project End
1996-06-30
Budget Start
1994-07-01
Budget End
1995-06-30
Support Year
4
Fiscal Year
1994
Total Cost
Indirect Cost
Name
University of Maryland Baltimore
Department
Microbiology/Immun/Virology
Type
Schools of Medicine
DUNS #
003255213
City
Baltimore
State
MD
Country
United States
Zip Code
21201
Jucker, M; McKenna, K; da Silva, A J et al. (1997) The Fes protein-tyrosine kinase phosphorylates a subset of macrophage proteins that are involved in cell adhesion and cell-cell signaling. J Biol Chem 272:2104-9
Jucker, M; Schiffer, C A; Feldman, R A (1997) A tyrosine-phosphorylated protein of 140 kD is constitutively associated with the phosphotyrosine binding domain of Shc and the SH3 domains of Grb2 in acute myeloid leukemia cells. Blood 89:2024-35
Izuhara, K; Feldman, R A; Greer, P et al. (1996) Interleukin-4 induces association of the c-fes proto-oncogene product with phosphatidylinositol-3 kinase. Blood 88:3910-8
Jucker, M; Feldman, R A (1995) Identification of a new adapter protein that may link the common beta subunit of the receptor for granulocyte/macrophage colony-stimulating factor, interleukin (IL)-3, and IL-5 to phosphatidylinositol 3-kinase. J Biol Chem 270:27817-22
Areces, L B; Dello Sbarba, P; Jucker, M et al. (1994) Functional specificity of cytoplasmic and transmembrane tyrosine kinases: identification of 130- and 75-kilodalton substrates of c-fps/fes tyrosine kinase in macrophages. Mol Cell Biol 14:4606-15
Izuhara, K; Feldman, R A; Greer, P et al. (1994) Interaction of the c-fes proto-oncogene product with the interleukin-4 receptor. J Biol Chem 269:18623-9
Areces, L B; Jucker, M; San Miguel, J A et al. (1993) Ligand-dependent transformation by the receptor for human granulocyte/macrophage colony-stimulating factor and tyrosine phosphorylation of the receptor beta subunit. Proc Natl Acad Sci U S A 90:3963-7