Involvement of integrins and Src in insulin signaling toward autophagic proteolysis in rat liver.
Cell volume changes critically determine hepatic signal transduction and metabolism. Hepatocyte swelling by insulin contributes to p38(MAPK) activation leading to inhibition of autophagic proteolysis. Recently integrins were shown to sense hypoosmotic hepatocyte swelling. Here the role of integrins, Src, and focal adhesion kinase (FAK) in insulin signaling was investigated using the intact organ model of perfused rat liver. Insulin increases [Tyr(P)(418)]Src, [Tyr(P)(397)]FAK, and dual p38(MAPK) phosphorylation by about 2-fold. Infusion of the integrin-antagonizing hexapeptide GRGDSP or the Src inhibitor PP-2 prevented activation of Src and p38(MAPK) and, consequently, proteolysis inhibition by insulin. However, insulin-induced phosphorylation of IRbeta (Tyr(1158)) and protein kinase B (PKB, Ser(473)), as well as K(+)-uptake and cell swelling, was not reduced by the inhibitors. Both hypoosmotic swelling and insulin increase the plasma membrane levels of activated beta(1) integrin. Inhibition of insulin-induced swelling by furosemide largely abolished activation of beta(1) integrin and phosphorylation of Src, but not of PKB. Rapamycin does not affect either insulin-induced K(+)-retention and cell swelling or proteolysis inhibition, indicating that swelling-dependent proteolysis inhibition occurs independently from the mammalian target of rapamycin. The data suggest that sensing of cell swelling by integrins essentially contributes to insulin signaling, thereby defining a novel way of integrin involvement in growth factor signaling.
Schliess
Freimut
F
Division of Gastroenterology, Hepatology, and Infectious Diseases, Heinrich Heine University, Moorenstrasse 5, D-40225 Düsseldorf, Germany. Freimut.Schliess@uni-duesseldorf.de
Reissmann
Regina
R
Reinehr
Roland
R
vom Dahl
Stephan
S
Häussinger
Dieter
D
D000818:Animals; D001343:Autophagy / Q000187:drug effects; D004232:Diuretics / Q000494:pharmacology; D007328:Insulin / Q000494:pharmacology*; D016023:Integrins / Q000502:physiology*; D007700:Kinetics; D008099:Liver / Q000187:drug effects / Q000502:physiology*; D008297:Male; D009842:Oligopeptides / Q000494:pharmacology; D010477:Perfusion; D016392:Proto-Oncogene Proteins pp60(c-src) / Q000037:antagonists & inhibitors / Q000502:physiology*; D051381:Rats; D017208:Rats, Wistar; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology*; D020123:Sirolimus / Q000494:pharmacology
D004232:Diuretics; D007328:Insulin; D016023:Integrins; D009842:Oligopeptides; C058161:glycyl-arginyl-glycyl-aspartyl-seryl-proline; C058347:glycyl-arginyl-glycyl-glutamyl-seryl-proline; D016392:Proto-Oncogene Proteins pp60(c-src); D020123:Sirolimus