The plecomacrolide vacuolar-ATPase inhibitor bafilomycin, alters insulin signaling in MIN6 beta-cells.
Inhibition of endosomal acidification disturbs insulin signaling in both liver and adipose cells. In this study we used MIN6 beta cells to determine whether bafilomycin, a potent inhibitor of the proton-translocating vacuolar ATPase, disrupts insulin signaling in islet beta cells. Pretreatment of MIN6 cells with varying concentrations of bafilomycin according to a time course revealed concentration and time-dependent changes in phosphorylation of insulin receptor signaling components. Increased phosphorylation of insulin receptor (IR), IRS2 and Akt was prolonged at low bafilomycin concentrations (10 and 50 nmol/L), whereas at high concentrations (100 and 200 nmol/L) phosphorylation rapidly returned to basal levels or below. Akt activation was demonstrated by transient increases in phosphorylation of BAD, cytoplasmic retention of FoxO1 and increased preproinsulin mRNA. Bcl2 expression was also transiently increased but reduced after 30 min exposure to bafilomycin, and this coincided with reduced cell viability. Thus, in beta cells inhibition of endosomal acidification by low concentrations of bafilomycin transiently increases insulin signaling, whereas high concentrations promote cell death. Bafilomycin and other agents that interfere with insulin signaling may contribute to diabetes development through disturbing homeostatic control of beta cell growth.
Department of Biochemistry and Molecular Biology, Monash University, Clayton, Victoria, Australia.
K D
KD
Hettiarachchi
P Z
PZ
Zimmet
M A
MA
Myers
D000143:Acids / Q000378:metabolism; D000818:Animals; D002460:Cell Line; D007328:Insulin / Q000378:metabolism*; D018942:Macrolides / Q000494:pharmacology*; D051379:Mice; D010766:Phosphorylation; D011384:Proinsulin / Q000378:metabolism; D011498:Protein Precursors / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism*; D015398:Signal Transduction; D025262:Vacuolar Proton-Translocating ATPases / Q000037:antagonists & inhibitors*; D014617:Vacuoles / Q000187:drug effects / Q000502:physiology; D051025:bcl-Associated Death Protein / Q000378:metabolism
D000143:Acids; C494820:Bad protein, mouse; D007328:Insulin; D018942:Macrolides; D011498:Protein Precursors; D012333:RNA, Messenger; D051025:bcl-Associated Death Protein; C013737:preproinsulin; C052501:bafilomycin B1; D011384:Proinsulin; D025262:Vacuolar Proton-Translocating ATPases