This proposal outlines studies to characterize the biosynthesis of the mononuclear phagocyte colony-stimulating factor CSF-1 (or M-CSF) and the interaction of this homodimeric growth factor with its receptor. The primary translation products of CSF-1 mRNAs are integral transmembrane proteins from which the soluble growth factor is derived by proteolytic cleavage. The products of two different human CSF-1 cDNAs will be compared with respect to the subcellular location and specificity of proteolytic processing. Site-directed mutagenesis will be used to define the extent of polypeptide sequences required for CSF-1 function and to identify cysteine residues that participate in disulfide bonds critical for dimer formation and biological activity. Membrane-bound CSF-1 molecules that fail to undergo proteolytic processing will be engineered and used to determine whether cell surface CSF-1 can directly stimulate receptor- bearing cells. The topological requirements of ligand-receptor interactions to induce autocrine transformation of cells simultaneously expressing CSF-1 and its receptor will also be investigated. These studies will be general interest to the biosynthesis of polypeptide growth factors and the mechanisms of growth factor-receptor interactions. A possible role for CSF-1 and its receptor outside the context of hematopoiesis will be examined by immunocytochemical techniques and in situ hybridization with specific molecular probes. An intensive study of the structure and expression of this growth factor is merited by the prospects for use of CSF-1 as a biological response modifier to stimulate macrophage production of function in clinical therapeutics.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
First Independent Research Support & Transition (FIRST) Awards (R29)
Project #
7R29HL040603-02
Application #
3472041
Study Section
Cellular Biology and Physiology Subcommittee 1 (CBY)
Project Start
1990-04-01
Project End
1994-03-31
Budget Start
1990-04-01
Budget End
1991-03-31
Support Year
2
Fiscal Year
1990
Total Cost
Indirect Cost
Name
Children's Hospital of Los Angeles
Department
Type
DUNS #
094878337
City
Los Angeles
State
CA
Country
United States
Zip Code
90027
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Deng, P; Wang, Y L; Pattengale, P K et al. (1996) The role of individual cysteine residues in the processing, structure, and function of human macrophage colony-stimulating factor. Biochem Biophys Res Commun 228:557-66
Umiel, T; Rettenmier, C W; Siegel, S et al. (1993) Establishment and characterization of a human mixed-lineage, T-lymphoid/myeloid cell line (USP-91). Blood 82:1829-37
Stein, J; Rettenmier, C W (1991) Proteolytic processing of a plasma membrane-bound precursor to human macrophage colony-stimulating factor (CSF-1) is accelerated by phorbol ester. Oncogene 6:601-5
Jackowski, S; Rettenmier, C W; Rock, C O (1990) Prostaglandin E2 inhibition of growth in a colony-stimulating factor 1-dependent macrophage cell line. J Biol Chem 265:6611-6
Stein, J; Borzillo, G V; Rettenmier, C W (1990) Direct stimulation of cells expressing receptors for macrophage colony-stimulating factor (CSF-1) by a plasma membrane-bound precursor of human CSF-1. Blood 76:1308-14
Rettenmier, C W (1989) Biosynthesis of macrophage colony-stimulating factor (CSF-1): differential processing of CSF-1 precursors suggests alternative mechanisms for stimulating CSF-1 receptors. Curr Top Microbiol Immunol 149:129-41
Rettenmier, C W; Sherr, C J (1989) The mononuclear phagocyte colony-stimulating factor (CSF-1, M-CSF). Hematol Oncol Clin North Am 3:479-93