Actions of PP2A on the MAP kinase pathway and apoptosis are mediated by distinct regulatory subunits.
Individual subunits of protein phosphatase 2A (PP2A), protein phosphatase 4, and protein phosphatase 5 were knocked out in Drosophila Schneider 2 cells by using RNA interference. Ablation of either the scaffold (A) or catalytic (C) subunits of PP2A caused the disappearance of all PP2A subunits. Treating cells with double-stranded RNA targeting all four of the Drosophila PP2A regulatory subunits caused the disappearance of both the A and C subunits. The loss of PP2A subunits was associated with decreased protein stability indicating that only the heterotrimeric forms of PP2A are stable in intact cells. Ablation of total PP2A by using double-stranded RNA against either the A or C subunit, or specific ablation of the R2/B regulatory subunit, enhanced insulin-induced ERK activation. These results indicated that the R2/B subunit targets PP2A to the mitogen-activated protein (MAP) kinase cascade in Schneider 2 cells, where it acts as a negative regulator. A severe loss of viability occurred in cells in which total PP2A or both isoforms of the Drosophila R5/B56 subunit had been ablated. The reduced viability of these cells correlated with the induction of markers of apoptosis including membrane blebbing and stimulation of caspase-3-like activity. These observations indicated that PP2A has a powerful antiapoptotic activity that is specifically mediated by the R5/B56 regulatory subunits. In contrast to PP2A, ablation of protein phosphatase 4 caused only a slight reduction in cell growth but had no effect on MAP kinase signaling or apoptosis. Depletion of protein phosphatase 5 had no effects on MAP kinase, cell growth, or apoptosis.
Silverstein
Adam M
AM
Department of Pharmacology, University of Texas Southwestern Medical Center, Dallas, TX 75390-9041, USA.
Barrow
Christina A
CA
Davis
Anthony J
AJ
Mumby
Marc C
MC
D000818:Animals; D017209:Apoptosis; D002478:Cells, Cultured; D004330:Drosophila; D004789:Enzyme Activation; D020935:MAP Kinase Signaling System / Q000502:physiology*; D010749:Phosphoprotein Phosphatases / Q000737:chemistry / Q000502:physiology*; D054648:Protein Phosphatase 2; D021122:Protein Subunits; D012330:RNA, Double-Stranded / Q000494:pharmacology
D021122:Protein Subunits; D012330:RNA, Double-Stranded; D010749:Phosphoprotein Phosphatases; D054648:Protein Phosphatase 2