title: AMP-activated Protein Kinase Suppresses Biosynthesis of Glucosylceramide by Reducing Intracellular Sugar Nucleotides.
abstract: The membrane glycolipid glucosylceramide (GlcCer) plays a critical role in cellular homeostasis. Its intracellular levels are thought to be tightly regulated. How cells regulate GlcCer levels remains to be clarified. AMP-activated protein kinase (AMPK), which is a crucial cellular energy sensor, regulates glucose and lipid metabolism to maintain energy homeostasis. Here, we investigated whether AMPK affects GlcCer metabolism. AMPK activators (5-aminoimidazole-4-carboxamide 1-β-d-ribofuranoside and metformin) decreased intracellular GlcCer levels and synthase activity in mouse fibroblasts. AMPK inhibitors or AMPK siRNA reversed these effects, suggesting that GlcCer synthesis is negatively regulated by an AMPK-dependent mechanism. Although AMPK did not affect the phosphorylation or expression of GlcCer synthase, the amount of UDP-glucose, an activated form of glucose required for GlcCer synthesis, decreased under AMPK-activating conditions. Importantly, the UDP-glucose pyrophosphatase Nudt14, which degrades UDP-glucose, generating UMP and glucose 1-phosphate, was phosphorylated and activated by AMPK. On the other hand, suppression of Nudt14 by siRNA had little effect on UDP-glucose levels, indicating that mammalian cells have an alternative UDP-glucose pyrophosphatase that mainly contributes to the reduction of UDP-glucose under AMPK-activating conditions. Because AMPK activators are capable of reducing GlcCer levels in cells from Gaucher disease patients, our findings suggest that reducing GlcCer through AMPK activation may lead to a new strategy for treating diseases caused by abnormal accumulation of GlcCer.
authors.affiliation: Laboratory for Molecular Membrane Neuroscience, RIKEN Brain Science Institute, Wako-shi, Saitama 351-0198, Japan.
authors.forename: Yohei
authors.initials: Y
authors.lastname: Ishibashi
authors.affiliation: Laboratory for Molecular Membrane Neuroscience, RIKEN Brain Science Institute, Wako-shi, Saitama 351-0198, Japan. Electronic address: hirabaya@riken.jp.
authors.forename: Yoshio
authors.initials: Y
authors.lastname: Hirabayashi
mesh_terms: D055372:AMP-Activated Protein Kinases / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D016466:CHO Cells; D002460:Cell Line; D006224:Cricetinae; D003412:Cricetulus; D004285:Dogs; D004789:Enzyme Activation; D005963:Glucosylceramides / Q000032:analysis / Q000378:metabolism*; D057809:HEK293 Cells; D006367:HeLa Cells; D006801:Humans; D051379:Mice; D041681:NIH 3T3 Cells; D010766:Phosphorylation; D011755:Pyrophosphatases / Q000235:genetics / Q000378:metabolism; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics; D014532:Uridine Diphosphate Glucose / Q000378:metabolism*
chemical_list: D005963:Glucosylceramides; D034741:RNA, Small Interfering; D055372:AMP-Activated Protein Kinases; D011755:Pyrophosphatases; C546204:Nudt14 protein, mouse; D014532:Uridine Diphosphate Glucose
keywords: AMP-activated kinase (AMPK); Gaucher disease; UDP-glucose pyrophosphatase; glucosylceramide; glycolipid; glycosphingolipid; lipid; metformin; sphingolipid; sugar nucleotide