S6 kinase p90rsk in granulocyte-macrophage colony-stimulating factor-stimulated proliferative and mature hematopoietic cells.

The ribosomal S6 kinase p90(rsk) was studied in mature and proliferating hemopoietic cells in response to the human cytokine granulocyte-macrophage colony-stimulating factor (GM-CSF). In neutrophils, GM-CSF induced time-dependent electrophoretic mobility shifts in immunoreactive p90(rsk). Although these shifts suggested changes in the phosphorylation status of the molecule, a kinase assay with whole cell lysates detected minimal (1.5-fold) increments in enzymatic activity. Only immunoprecipitation followed by immune complex kinase assay or in-gel kinase assay performed against the RSK substrate RRLSSLRA evidenced an increase in p90(rsk) activity (3.4-fold). p90(rsk) was also detected in the GM-CSF-dependent erythroleukemia cell line TF-1. Normally cultured, cytokine-supplemented cells did not respond to further GM-CSF stimulation. However, the activity of p90(rsk) in cytokine-starved cells increased dramatically in response to short term GM-CSF challenge. This effect was readily observable in total cell lysates (6.6-fold increase over controls) and was paralleled by changes in mitogen-activated protein kinase activity (a substrate of p90(rsk)). Thus, p90(rsk) is present in mature hemopoietic cells, but the extent of the enzymatic response to GM-CSF is significantly lower than that seen in proliferative cells.