title: Spermine increases phosphatidylinositol 4,5-bisphosphate content in permeabilized and nonpermeabilized HL60 cells.
abstract: The polyamine spermine (N,N'bis[3-aminopropyl]-1,4-butanediamine) activates phosphatidylinositol-4-phosphate 5-kinase (PtdIns(4)P5K) and phosphatidylinositol 4-kinase (PtdIns4K) in vitro. Spermine concentration increases that occur in proliferating cells were approximated in streptolysin O-permeabilized HL60 cells. When phospholipase C was activated by GTPgammaS in the presence of PITPalpha, 0.1-1.2 mM spermine evoked increases in PtdIns(4,5)P(2) contents in a dose-dependent manner to 110-170% of control and concomitantly decreased inositol phosphate formation by 10-50%. Spermine-induced increases in PtdIns(4,5)P(2) content in permeabilized cells also occurred during GTPgammaS stimulation in the absence of PITPalpha, were augmented in the presence of PITPalpha, occurred in unstimulated cells and were additive to PtdIns(4,5)P(2) formation evoked by ARF1, another activator of phosphoinositide kinases. Slowly developing spermine-evoked increases in PtdIns(4,5)P(2) contents occurred in nonpermeabilized cells that were abolished in the presence of a spermine transport inhibitor. Data are consistent with spermine at physiological concentrations evoking a PITPalpha-dependent shift in formation of PtdIns(4,5)P(2) from compartments that contained an active phospholipase C to compartments that were separated from an active PLC and from PtdIns(4,5)P(2) formed by ARF1.
authors.affiliation: Department of Physiology, University of Pennsylvania, School of Medicine, Philadelphia, PA 19104, USA. rfcoburn@mail.med.upenn.edu
authors.forename: Ronald F
authors.initials: RF
authors.lastname: Coburn
authors.forename: David H
authors.initials: DH
authors.lastname: Jones
authors.forename: Clive P
authors.initials: CP
authors.lastname: Morgan
authors.forename: Carl B
authors.initials: CB
authors.lastname: Baron
authors.forename: Shamshad
authors.initials: S
authors.lastname: Cockcroft
mesh_terms: D020823:ADP-Ribosylation Factor 1 / Q000494:pharmacology; D001426:Bacterial Proteins; D004305:Dose-Response Relationship, Drug; D004789:Enzyme Activation / Q000187:drug effects; D016244:Guanosine 5'-O-(3-Thiotriphosphate); D018922:HL-60 Cells / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D007295:Inositol Phosphates / Q000032:analysis / Q000378:metabolism; D019269:Phosphatidylinositol 4,5-Diphosphate / Q000032:analysis / Q000378:metabolism*; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000378:metabolism; D013096:Spermine / Q000037:antagonists & inhibitors / Q000494:pharmacology*; D013301:Streptolysins; D013997:Time Factors
chemical_list: D001426:Bacterial Proteins; D007295:Inositol Phosphates; D019269:Phosphatidylinositol 4,5-Diphosphate; D013301:Streptolysins; C025964:streptolysin O; D013096:Spermine; D016244:Guanosine 5'-O-(3-Thiotriphosphate); D017853:Phosphotransferases (Alcohol Group Acceptor); C068961:phosphatidylinositol 4,5-biphosphate kinase; D020823:ADP-Ribosylation Factor 1