Phospholipase D1 regulates high-affinity IgE receptor-induced mast cell degranulation.
To investigate the role of phospholipase D (PLD) in FcepsilonRI signaling, the wild-type or the catalytically inactive forms of PLD1 or PLD2 were stably overexpressed in RBL-2H3 mast cells. FcepsilonRI stimulation resulted in the activation of both PLD1 and PLD2. However, PLD1 was the source of most of the receptor-induced PLD activity. There was enhanced FcepsilonRI-induced degranulation only in cells that overexpressed the catalytically inactive PLD1. This dominant-negative PLD1 enhanced FcepsilonRI-induced tyrosine phosphorylations of early signaling molecules such as the receptor subunits, Syk and phospholipase C-gamma which resulted in faster release of Ca(2+) from intracellular sources. Therefore, PLD1 negatively regulates signals upstream of the Ca(2+) response. However, FcepsilonRI-induced PLD activation required Syk and was downstream of the Ca(2+)response, suggesting that basal PLD1 activity rather than that activated by cell stimulation controlled these early signaling events. Dominant-negative PLD1 reduced the basal phosphatidic acid formation in unstimulated cells, which was accompanied by an increase in FcepsilonRI within the lipid rafts. These results indicate that constitutive basal PLD1 activity by regulating phosphatidic acid formation controls the early signals initiated by FcepsilonRI aggregation that lead to mast cell degranulation.
Hitomi
Tomohiro
T
Receptors and Signal Transduction Section, Oral Infection and Immunity Branch, National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, MD 20892, USA. thitomi@mail.nih.gov
Zhang
Juan
J
Nicoletti
Liliana M
LM
Grodzki
Ana Cristina G
AC
Jamur
Maria C
MC
Oliver
Constance
C
Siraganian
Reuben P
RP
D000818:Animals; D002118:Calcium / Q000502:physiology; D002458:Cell Fractionation; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000276:immunology; D003594:Cytoplasmic Granules / Q000276:immunology*; D008407:Mast Cells / Q000276:immunology*; D010739:Phospholipase D / Q000378:metabolism*; D010766:Phosphorylation; D019000:Phosphotyrosine / Q000378:metabolism; D051381:Rats; D017455:Receptors, IgE / Q000276:immunology*
D017455:Receptors, IgE; D019000:Phosphotyrosine; C105606:phospholipase D2; D010739:Phospholipase D; C106801:phospholipase D1; D002118:Calcium