Identification and characterization of a nucleotide binding site on recombinant murine granulocyte/macrophage-colony stimulating factor

1992 ◽  
Vol 3 (6) ◽  
pp. 484-492 ◽  
Author(s):  
Michael A. Doukas ◽  
Ashok J. Chavan ◽  
Cecelia Gass ◽  
Thomas Boone ◽  
Boyd E. Haley
Keyword(s):  
Binding Site ◽  
Nucleotide Binding ◽  
Nucleotide Binding Site ◽  
Colony Stimulating Factor ◽  
Macrophage Colony Stimulating Factor ◽  
Macrophage Colony ◽  
Identification And Characterization ◽  
Stimulating Factor

scholarly journals Granulocyte-macrophage colony-stimulating factor and interleukin-3 signaling pathways converge on the CREB-binding site in the human egr-1 promoter.

1994 ◽  
Vol 14 (9) ◽  
pp. 5975-5985 ◽  
Author(s):  
K M Sakamoto ◽  
J K Fraser ◽  
H J Lee ◽  
E Lehman ◽  
J C Gasson
Keyword(s):  
Binding Site ◽  
Electrophoretic Mobility ◽  
Colony Stimulating Factor ◽  
Mobility Shift ◽  
Response Gene ◽  
Gm Csf ◽  
Macrophage Colony Stimulating Factor ◽  
Macrophage Colony ◽  
Electrophoretic Mobility Shift Assays ◽  
Stimulating Factor

Granulocyte-macrophage colony-stimulating factor (GM-CSF) stimulates myeloid progenitor cell proliferation and enhances the function of terminally differentiated effector cells. Interleukin-3 (IL-3) stimulation results in the proliferation and maturation of early bone marrow progenitor cells. These activities are mediated by non-tyrosine kinase-containing receptors which consist of ligand-specific alpha subunits that complex with a common beta subunit required for signal transduction. Both GM-CSF and IL-3 rapidly and transiently induce expression of early growth response gene 1 (egr-1) in the human factor-dependent cell line TF-1. To define the mechanism of early response gene induction by GM-CSF and IL-3, growth factor- and serum-starved TF-1 cells transfected with recombinant constructs containing sequences of the human egr-1 promoter were stimulated with GM-CSF or IL-3. A 116-nucleotide (nt) region of the egr-1 promoter which contains sequences inducible by GM-CSF and IL-3 was defined. DNase I footprint analysis identified a 20-nt region, including nt -57 to -76, which contains a potential cyclic AMP (cAMP) response element (CRE). Electrophoretic mobility shift assays performed with CREB antibody confirmed the presence of CREB in the DNA-binding complex. Mutational analysis of the cytokine-responsive region of the egr-1 promoter revealed that both the cAMP response and serum response elements are required for induction by GM-CSF and IL-3. Nuclear extracts from GM-CSF- or IL-3-stimulated but not unstimulated TF-1 cells contain factors which specifically bind to the Egr-1-binding site in the nt -600 to -480 region of the promoter. Electrophoretic mobility shift assays were performed with antibodies against the Egr-1 protein to demonstrate the presence of the protein product in the shifted complex. Our studies suggest that the Egr-1 protein may further stimulate transcription of the egr-1 gene in response to GM-CSF as a secondary event.

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