Activation and mitochondrial translocation of protein kinase C delta are necessary for insulin stimulation of pyruvate dehydrogenase complex activityin muscle and liver cells

In L6 skeletal muscle cells and immortalized hepatocytes, insulin induced a 2-fold increase in the activity of the pyruvate dehydrogenase (PDH) comple x. This effect was almost completely blocked by the protein kinase C (PKC) delta inhibitor Rottlerin and by PKC delta antisense oligonucleotides. At v ariance, overexpression of wild-type PKC delta or of an active PKC delta mu tant induced PDH complex activity in both L6 and liver cells. Insulin stimu lation of the activity of the PDH complex was accompanied by a 2.5-fold inc rease in PDH phosphatases 1 and 2 (PDP1/2) activity with no change in the a ctivity of PDH kinase. PKC delta antisense blocked insulin activation of PD P1/2, the same as with PDH. In insulin-exposed cells, PDP1/2 activation was paralleled by activation and mitochondrial translocation of PKC8 delta, as revealed by cell subfractionation and confocal microscopy studies. The mit ochondrial translocation of PKC delta, like its activation, was prevented b y Rottlerin. In extracts from insulin-stimulated cells, PKC delta co-precip itated with PDP1/2. PKC delta also bound to PDP1/2 in overlay blots, sugges ting that direct PKC delta -PDP interaction may occur in vivo as well. In i ntact cells, insulin exposure determined PDP1/2 phosphorylation, which was specifically prevented by PKC delta antisense. PKC delta also phosphorylate d PDP in vitro, followed by PDP1/2 activation. Thus, in muscle and liver ce lls, insulin causes activation and mitochondrial translocation of PKC delta , accompanied by PDP phosphorylation and activation. These events are neces sary for insulin activation of the PDH complex in these cells.