Functional Differentiation Signals Mediated by Distinct Regions of the Cytoplasmic Domain of the Granulocyte Colony-Stimulating Factor Receptor

Abstract

Granulocyte colony-stimulating factor receptor (G-CSFR) regulates the proliferation and differentiation of neutrophilic progenitor cells through interaction with its cytokine. Exposure of WEHI-3B D+ myelomonocytic leukemia and myeloid LGM-1 cells overexpressing the G-CSFR to G-CSF resulted in induction of differentiation as measured by (1) the ability to reduce nitroblue tetrazolium (NBT), (2) the expression of Mac-I antigen, and (3) the expression of FcγII/III receptor. Mutational analyses indicated that distinct regions of the cytoplasmic domain were critical for efficient induction of each functional marker. The membrane proximal region containing homology sequences of boxes 1 and 2 was important for the activation of all three functional markers of mature neutrophils. Induction of the capacities to express Mac-I antigen or FcγII/III receptor also required additional sequences in the membrane proximal region between amino acids 70 and 100 and may be dependent on the phosphorylation of Tyr703. The findings suggest that distinct sequences within the amino-terminal region of the cytoplasmic domain of the receptor are sufficient to induce these functional markers of differentiation, and receptor tyrosine phosphorylation may be necessary.