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Internalization of the granulocyte-macrophage colony-stimulating factor
receptor is not required for induction of protein tyrosine phosphorylation
in human myeloid cells
K Okuda, B Druker, Y Kanakura, M Koenigsmann and JD Griffin
Division of Tumor Immunology, Dana-Farber Cancer Institute, Boston, MA
02115.
Granulocyte-macrophage colony-stimulating factor (GM-CSF) exerts its
biologic activities through binding to specific high-affinity cell surface
receptors. After binding, the ligand/receptor complex is rapidly
internalized in most hematopoietic cells. Using a human factor- dependent
cell line, MO7, and normal human neutrophils, we found that the
internalization is exquisitely temperature-dependent, such that
ligand/receptor internalization does not detectably occur at 4 degrees C.
Activation of the GM-CSF receptor has previously been shown to stimulate a
number of postreceptor signal transduction pathways, including activation
of a tyrosine kinase and activation of the serine/threonine kinase, Raf-1.
The GM-CSF-stimulated increase in tyrosine kinase activity occurs rapidly
at both 4 degrees C and 37 degrees C, and therefore is likely to be
independent of receptor internalization. At 37 degrees C, the protein
tyrosine phosphorylation was transient in MO7 cells, with maximum
phosphorylation observed after 5 to 15 minutes, followed by a rapid
decline. At 4 degrees C, the protein tyrosine phosphorylation of the same
substrates was greater than at 37 degrees C, and no decline in substrate
phosphorylation was observed for at least 90 minutes. In contrast to
tyrosine phosphorylation, the activation and hyper-phosphorylation of Raf-1
observed at 37 degrees C in both MO7 cells and neutrophils was markedly
diminished at 4 degrees C. These results indicate that at least one
postreceptor signal transduction mechanism, activation of a tyrosine
kinase, does not require ligand/receptor internalization, and indicate that
receptor internalization may be a consequence, rather than the initiator,
of signal transduction.
Volume 78,
Issue 8,
pp. 1928-1935,
10/15/1991
Copyright © 1991 by The American Society of Hematology
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