Phosphatidylinositol 3-kinase activity is required for early endosome fusion.
The homotypic fusion between early endosomes from baby-hamster kidney cells is blocked by addition of the fungal metabolite wortmannin with an IC50 of approx. 15 nM. Over this concentration range, wortmannin has been regarded as a specific inhibitor of phosphatidylinositol (PI) 3-kinase. Further confirmation of the participation of a PI 3-kinase in the fusion reaction has been obtained by demonstrating a sensitivity to an additional, structurally unrelated, PI 3-kinase inhibitor, LY294002 [2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one]. Assays constructed such that only the membranous component has been incubated with wortmannin show in vitro fusion to be sensitive to treatment with the drug. Assays in which only the cytosolic component has been treated with wortmannin also showed inhibition of in vitro fusion, but to a lesser extent. PI 3-kinase action almost certainly involves direct regulation of membrane fusion, as no vesicular intermediate has been identified, despite previous extensive morphological examination of in vitro endosome fusions.
Jones
A T
AT
Physiological Laboratory, University of Liverpool, U.K.
Clague
M J
MJ
D000730:Androstadienes / Q000494:pharmacology; D000818:Animals; D015153:Blotting, Western; D002417:Cattle; D002460:Cell Line; D002867:Chromones / Q000494:pharmacology; D006224:Cricetinae; D011992:Endosomes / Q000648:ultrastructure*; D051275:Glucose Transporter Type 4; D007425:Intracellular Membranes / Q000502:physiology*; D007668:Kidney; D008099:Liver / Q000648:ultrastructure; D008561:Membrane Fusion / Q000187:drug effects; D009004:Monosaccharide Transport Proteins / Q000378:metabolism; D009025:Morpholines / Q000494:pharmacology; D009124:Muscle Proteins; D019869:Phosphatidylinositol 3-Kinases; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000037:antagonists & inhibitors / Q000378:metabolism*; D000077191:Wortmannin
D000730:Androstadienes; D002867:Chromones; D051275:Glucose Transporter Type 4; D009004:Monosaccharide Transport Proteins; D009025:Morpholines; D009124:Muscle Proteins; C085911:2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one; D019869:Phosphatidylinositol 3-Kinases; D017853:Phosphotransferases (Alcohol Group Acceptor); D000077191:Wortmannin