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Plant Physiol, August 2001, Vol. 126, pp. 1507-1518
Plasma Membrane Phosphatidylinositol 4,5-Bisphosphate Levels
Decrease with Time in Culture1
Ingo
Heilmann,
Imara Y.
Perera,
Wolfgang
Gross, and
Wendy F.
Boss*
Department of Botany, North Carolina State University, Raleigh, NC
27695-7612 (I.H., I.Y.P., W.F.B.); and Institut für
Biologie (Pflanzenphysiologie), Freie Universität, Berlin,
Germany (W.G.)
During the stationary phase of growth, after 7 to 12 d in culture,
the levels of phosphatidylinositol 4,5-bisphosphate
(PtdInsP2) decreased by 75% in plasma membranes of the red
alga Galdieria sulphuraria. Concomitant with the
decrease in PtdInsP2 levels in plasma membranes, there was
an increase in PtdInsP2 in microsomes, suggesting that the
levels of plasma membrane PtdInsP2 are regulated differentially. The decline of PtdInsP2 in plasma membranes
was accompanied by a 70% decrease in the specific activity of PtdInsP kinase and by reduced levels of protein cross-reacting with antisera against a conserved PtdInsP kinase domain. Upon osmotic stimulation, the loss of PtdInsP2from the plasma membrane increased from
10% in 7-d-old cells to 60% in 12-d-old cells, although the levels of
inositol 1,4,5-trisphosphate (InsP3) produced in whole
cells were roughly equal at both times. When cells with low plasma
membrane PtdInsP2 levels were osmotically stimulated, a
mild osmotic stress (12.5 mM KCl) activated PtdInsP kinase
prior to InsP3 production, whereas in cells with high
plasma membrane PtdInsP2, more severe stress (250 mM KCl) was required to induce an increase in PtdInsP kinase activity. The differential regulation of a plasma membrane signaling pool of PtdInsP2 is discussed with regard to the
implications for understanding the responsive state of cells.
1
This work was supported by the National Science
Foundation (grant no. MCB-9604285 to W.F.B.) and by a Deutscher
Akademischer Austauschdienst fellowship (HSPIII to I.H.)
financed by the German Federal Ministry of Education, Science,
Research, and Technology.
*
Corresponding author; email wendy_boss{at}ncsu.edu; fax
919-515-3436.
© 2001 American Society of Plant Physiologists
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