Phospholipase D controls Dictyostelium development by regulating G protein signaling.
Dictyostelium discoideum cells normally exist as individual amoebae, but will enter a period of multicellular development upon starvation. The initial stages of development involve the aggregation of individual cells, using cAMP as a chemoattractant. Chemotaxis is initiated when cAMP binds to its receptor, cAR1, and activates the associated G protein, Gα2βγ. However, chemotaxis will not occur unless there is a high density of starving cells present, as measured by high levels of the secreted quorum sensing molecule, CMF. We previously demonstrated that cells lacking PldB bypass the need for CMF and can aggregate at low cell density, whereas cells overexpressing pldB do not aggregate even at high cell density. Here, we found that PldB controlled both cAMP chemotaxis and cell sorting. PldB was also required by CMF to regulate G protein signaling. Specifically, CMF used PldB, to regulate the dissociation of Gα2 from Gβγ. Using fluorescence resonance energy transfer (FRET), we found that along with cAMP, CMF increased the dissociation of the G protein. In fact, CMF augmented the dissociation induced by cAMP. This augmentation was lost in cells lacking PldB. PldB appears to mediate the CMF signal through the production of phosphatidic acid, as exogenously added phosphatidic acid phenocopies overexpression of pldB. These results suggest that phospholipase D activity is required for CMF to alter the kinetics of cAMP-induced G protein signaling.
Department of Biological Sciences, Hunter College, New York, New York 10065, USA.
Sibnath
S
Ray
Yi
Y
Chen
Joanna
J
Ayoung
Rachel
R
Hanna
Derrick
D
Brazill
D002633:Chemotaxis; D000242:Cyclic AMP / Q000502:physiology; D004023:Dictyostelium / Q000201:enzymology / Q000254:growth & development*; D020555:GTP-Binding Protein Regulators / Q000502:physiology; D019204:GTP-Binding Proteins / Q000502:physiology*; D010712:Phosphatidic Acids / Q000096:biosynthesis; D010739:Phospholipase D / Q000502:physiology*; D053038:Quorum Sensing; D011953:Receptors, Cyclic AMP / Q000378:metabolism; D015398:Signal Transduction
D020555:GTP-Binding Protein Regulators; D010712:Phosphatidic Acids; D011953:Receptors, Cyclic AMP; C068125:cyclic AMP receptor cAR1; D000242:Cyclic AMP; D010739:Phospholipase D; D019204:GTP-Binding Proteins