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- 2015.1
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AMP-activated protein kinase (AMPK) is a metabolic sensor activated during metabolic stress and it regulates various enzymes and cellular processes to maintain metabolic homeostasis. We previously reported that activation of AMPK by glucose deprivation (GD) and leptin increases KATP currents by increasing the surface levels of KATP channel proteins in pancreatic β-cells. Here, we show that the signaling mechanisms that mediate actin cytoskeleton remodeling are closely associated with AMPK-induced KATP channel trafficking. Using F-actin staining with Alexa 633-conjugated phalloidin, we observed that dense cortical actin filaments present in INS-1 cells cultured in 11mM glucose were disrupted by GD or leptin treatment. These changes were blocked by inhibiting AMPK using compound C or siAMPK and mimicked by activating AMPK using AICAR, indicating that cytoskeletal remodeling induced by GD or leptin was mediated by AMPK signaling. AMPK activation led to the activation of Rac GTPase and the phosphorylation of myosin regulatory light chain (MRLC). AMPK-dependent actin remodeling induced by GD or leptin was abolished by the inhibition of Rac with a Rac inhibitor (NSC23766), siRac1 or siRac2, and by inhibition of myosin II with a myosin ATPase inhibitor (blebbistatin). Immunocytochemistry, surface biotinylation and electrophysiological analyses of KATP channel activity and membrane potentials revealed that AMPK-dependent KATP channel trafficking to the plasma membrane was also inhibited by NSC23766 or blebbistatin. Taken together, these results indicate that AMPK/Rac-dependent cytoskeletal remodeling associated with myosin II motor function promotes the translocation of KATP channels to the plasma membrane in pancreatic β-cells.
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참고문헌 (22)
참고문헌 신청
Asahara S / 2013 / Ras-related C3 botulinum toxin substrate 1(RAC1)regulates glucose-stimulated insulin secretion via modulation of F-actin / Diabetologia 56 : 1088 ~ 1097
Blume C / 2007 / AMP-activated protein kinase impairs endothelial actin cytoskeleton assembly by phosphorylating vasodilator-stimulated phosphoprotein / J Biol Chem 282 : 460
Chen PC / 2013 / Leptin regulates KATP channel trafficking in pancreatic β-cells by a signaling mechanism involving AMP-activated protein kinase(AMPK)and cAMP-dependent pot
Dean SO / 2005 / Distinct pathways control recruitment and maintenance of myosin ll at the cleavage furrow during cytokinesis / Proc Natl Acad Sci USA 102 : 13473 ~ 13478
Ikonen E / 1997 / Myosin ll is associated with Golgi membranes : identification of p200 as nonmuscle myosin ll on Golgi-derived vesicles / J Cell Sci 110 : 2155 ~ 2164
Blume C / 2007 / AMP-activated protein kinase impairs endothelial actin cytoskeleton assembly by phosphorylating vasodilator-stimulated phosphoprotein / J Biol Chem 282 : 460
Chen PC / 2013 / Leptin regulates KATP channel trafficking in pancreatic β-cells by a signaling mechanism involving AMP-activated protein kinase(AMPK)and cAMP-dependent pot
Dean SO / 2005 / Distinct pathways control recruitment and maintenance of myosin ll at the cleavage furrow during cytokinesis / Proc Natl Acad Sci USA 102 : 13473 ~ 13478
Ikonen E / 1997 / Myosin ll is associated with Golgi membranes : identification of p200 as nonmuscle myosin ll on Golgi-derived vesicles / J Cell Sci 110 : 2155 ~ 2164
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