Ouabain affects cell migration via Na,K-ATPase-p130cas and via nucleus-centrosome association

• Published in: PloS one Vol. 12; no. 8; p. e0183343
• Main Authors: Ou, Young, Pan, Chen Xuan, Zuo, Jeremy, van der Hoorn, Frans A
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 17.08.2017; Public Library of Science (PLoS)
• Subjects: Amino Acid Sequence; Binding sites; Biochemistry; Biology and Life Sciences; Cell adhesion & migration; Cell growth; Cell Line; Cell migration; Cell Movement - drug effects; Cell Nucleus - drug effects; Centrosome; Centrosome - drug effects; Crk-Associated Substrate Protein - chemistry; Crk-Associated Substrate Protein - drug effects; Crk-Associated Substrate Protein - metabolism; Dephosphorylation; Dosage and administration; Enzymes; Extracellular signal-regulated kinase; Humans; Inhibitors; Kinases; Medicine; Medicine and Health Sciences; Membrane proteins; Molecular biology; Monoclonal antibodies; Na+/K+-exchanging ATPase; Nuclei; Nuclei (cytology); Ouabain; Ouabain - pharmacology; Phosphorylation; Potassium; Proteins; Research and Analysis Methods; Rodents; Signal transduction; Signaling; siRNA; Sodium; Sodium-Potassium-Exchanging ATPase - drug effects; Sodium-Potassium-Exchanging ATPase - metabolism; Src protein; Steroid hormones; Steroids; Canada; Calgary Alberta Canada
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0183343

Na,K-ATPase is a membrane protein that catalyzes ATP to maintain transmembrane sodium and potassium gradients. In addition, Na,K-ATPase also acts as a signal-transducing receptor for cardiotonic steroids such as ouabain and activates a number of signalling pathways. Several studies report that ouabain affects cell migration. Here we used ouabain at concentrations far below those required to block Na,K-ATPase pump activity and show that it significantly reduced RPE cell migration through two mechanisms. It causes dephosphorylation of a 130 kD protein, which we identify as p130cas. Src is involved, because Src inhibitors, but not inhibitors of other kinases tested, caused a similar reduction in p130cas phosphorylation and ouabain increased the association of Na,K-ATPase and Src. Knockdown of p130cas by siRNA reduced cell migration. Unexpectedly, ouabain induced separation of nucleus and centrosome, also leading to a block in cell migration. Inhibitor and siRNA experiments show that this effect is mediated by ERK1,2. This is the first report showing that ouabain can regulate cell migration by affecting nucleus-centrosome association.