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Vol. 58, Issue 3, 491-497, September 2000
Institut für Pharmakologie, Universitätsklinikum Essen,
Essen, Germany
Sphingosine-1-phosphate (SPP), produced by sphingosine kinase, has
recently been reported to act as an intracellular second messenger for
Ca2+ and mitogenic responses triggered by membrane
receptors and as an extracellular ligand for specific SPP receptors.
Here, we investigated the signaling pathway leading to SPP production
by the G protein-coupled P2Y2 receptor and its functional
implication in human leukemia (HL-60) cells, which do not respond to
extracellular SPP. P2Y2 receptor activation by UTP or ATP
resulted in rapid and transient production of SPP, which was
insensitive to pertussis toxin and blocked by the sphingosine kinase
inhibitor, DL-threo-dihydrosphingosine. Treatment of HL-60 cells with this inhibitor did not affect activation of mitogen-activated protein kinases, but suppressed
Ca2+ mobilization by the P2Y2 receptor.
However, receptor-induced SPP production apparently required an
increase in intracellular Ca2+ concentration, but not
Ca2+ influx, and was mimicked by exposure of cells to
Ca2+ ionophores. Taken together, activation of the
P2Y2 receptor stimulates SPP production in HL-60 cells, a
process apparently not required for mitogen-activated protein kinase
activation, but most likely representing an amplification system for
receptor-mediated Ca2+ signaling.
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