title: Lysophosphatidic acid-stimulated phosphorylation of PKD2 is mediated by PI3K p110β and PKCδ in myoblasts.
abstract: BACKGROUND
  G-protein coupled receptor (GPCR) signaling in skeletal muscle is incompletely understood; in particular, the signaling pathways that regulate GPCR-mediated signaling in skeletal muscle are only beginning to be established. Lysophosphatidic acid (LPA) is a GPCR agonist that has previously been shown to activate protein kinase D (PKD) in non-muscle cells; however, whether PKD is activated in response to LPA in skeletal muscle myoblasts, and the identities of signaling intermediates that regulate this activation, have not been defined.
  
  
  OBJECTIVE
  To determine whether PKD is activated in response to LPA administration in myoblasts, and to define the signaling pathways that mediate LPA-stimulated PKD phosphorylation.
  
  
  METHODS
  C2C12 myoblasts were treated with LPA and signaling pathways examined by means of Western immunoblotting and real-time PCR (RT-PCR). Pharmacological inhibition and RNA-interference were used to target specific molecules to determine their involvement in LPA-induced PKD phosphorylation.
  
  
  RESULTS
  Treatment of myoblasts with exogenous LPA revealed that PI3K p110β mediated PKD phosphorylation at Ser 748 and at Ser 916 through kinase-dependent and kinase-independent mechanisms. Loss of PKCδ, but not the loss of PKCα, prevented LPA-induced PKD phosphorylation. The PKD isoform responsive to LPA treatment was identified as PKD2.
  
  
  CONCLUSIONS
  These results indicate that LPA-stimulated PKD2 phosphorylation requires PKCδ and non-catalytic actions of PI3K p110β, and provide new information with respect to GPCR-mediated signal transduction in myoblasts.
authors.affiliation: US Army Research Institute of Environmental Medicine, Military Performance Division, Natick, MA 01760, USA.
authors.forename: Christine M
authors.initials: CM
authors.lastname: Lynch
authors.forename: Luis A
authors.initials: LA
authors.lastname: Leandry
authors.forename: Ronald W
authors.initials: RW
authors.lastname: Matheny
mesh_terms: D000818:Animals; D002460:Cell Line; D008246:Lysophospholipids / Q000494:pharmacology*; D051379:Mice; D032446:Myoblasts / Q000187:drug effects / Q000378:metabolism*; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism*; D010766:Phosphorylation / Q000187:drug effects; D051745:Protein Kinase C-delta / Q000378:metabolism*; D050396:TRPP Cation Channels / Q000378:metabolism*
chemical_list: D008246:Lysophospholipids; D019869:Phosphatidylinositol 3-Kinases; D051745:Protein Kinase C-delta; D050396:TRPP Cation Channels; C497259:Prkcd protein, mouse; C032881:lysophosphatidic acid