Protein kinase Cζ-dependent LKB1 serine 428 phosphorylation increases LKB1 nucleus export and apoptosis in endothelial cells (Publication with Expression of Concern. See vol. 294, pg. 10026, 2019) (Withdrawn Publication. See vol. 294, pg. 13831, 2019)

LKB1 is a serine-threonine protein kinase that, when inhibited, may result in unregulated cell growth and tumor formation. However, how LKB1 is regulated remains poorly understood. The aim of the present study was to define the upstream signaling events responsible for peroxynitrite (ONOO-)-induced LKB1 activation. Exposure of cultured human umbilical vein endothelial cells to a low concentration of ONOO- (5 mu M) significantly increased the phosphorylation of LKB1 at Ser(428) and protein kinase C zeta (PKC zeta) at Thr(410). These effects were accompanied by increased activity of the lipid phosphatase PTEN, decreased activity and phosphorylation (Ser(473)) of Akt, and induction of apoptosis. ONOO- enhanced Akt-Ser(473) phosphorylation in LKB1-deficient HeLa S3 cells or in HeLa S3 cells transfected with kinase-dead LKB1. Conversely, ONOO- inhibited Akt Ser(473) phosphorylation when wild type LKB1 were reintroduced in HeLa S3 cells. Further analysis revealed that PKC zeta directly phosphorylated LKB1 at Ser(428) in vitro and in intact cells, resulting in increased PTEN phosphorylation at Ser(380)/Thr(382)/ (383). Finally, ONOO- enhanced PKC zeta nuclear import and LKB1 nuclear export. We conclude that PKC zeta mediates LKB1-dependent Akt inhibition in response to ONOO-, resulting in endothelial apoptosis.