title: Hydrogen peroxide depletes phosphatidylinositol-3-phosphate from endosomes in a p38 MAPK-dependent manner and perturbs endocytosis.
abstract: Phosphatidylinositol-3-phosphate (PI3P) is a lipid that is enriched specifically in early endosomes. Given that early endosomes containing PI3P act as a microdomain to recruit proteins that contain a PI3P-binding domain (FYVE domain), the equilibrium between the production and degradation of PI3P influences a variety of processes, including endocytosis and signal transduction via endosomes. In the study reported herein, we have developed a novel analytical method to quantify the amount of PI3P in endosomes by introducing a GST-2xFYVE protein probe into semi-intact cells. The GST-2xFYVE probe was targeted specifically to intracellular PI3P-containing endosomes, which retained their small punctate structure, and allowed the semi-quantitative measurement of intracellular PI3P. Using the method, we found that treatment of HeLa cells with H(2)O(2) decreased the amount of PI3P in endosomes in a p38 MAPK-dependent manner. In addition, H(2)O(2) treatment delayed transport through various endocytic pathways, especially post-early endosome transport; the retrograde transport of cholera toxin was especially dependent on the amount of PI3P in endosomes. This article is part of a Special Issue entitled: 11th European Symposium on Calcium.
authors.affiliation: Department of Life Sciences, Graduate School of Arts and Sciences, The University of Tokyo, Komaba 3-8-1, Meguro-ku, Tokyo 153-8902, Japan.
authors.forename: Fumi
authors.initials: F
authors.lastname: Kano
authors.forename: Tamaki
authors.initials: T
authors.lastname: Arai
authors.forename: Mariko
authors.initials: M
authors.lastname: Matsuto
authors.forename: Hanako
authors.initials: H
authors.lastname: Hayashi
authors.forename: Moritoshi
authors.initials: M
authors.lastname: Sato
authors.forename: Masayuki
authors.initials: M
authors.lastname: Murata
mesh_terms: D001426:Bacterial Proteins / Q000378:metabolism; D001681:Biological Assay; D001692:Biological Transport / Q000187:drug effects; D002772:Cholera Toxin / Q000378:metabolism; D058546:Class III Phosphatidylinositol 3-Kinases / Q000378:metabolism; D004705:Endocytosis / Q000187:drug effects*; D011992:Endosomes / Q000187:drug effects* / Q000201:enzymology*; D066246:ErbB Receptors / Q000378:metabolism; D055785:Gene Knockdown Techniques; D006056:Golgi Apparatus / Q000187:drug effects / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology*; D007093:Imidazoles / Q000494:pharmacology; D047908:Intracellular Signaling Peptides and Proteins / Q000737:chemistry / Q000378:metabolism; D042541:Intracellular Space / Q000187:drug effects / Q000378:metabolism; D008164:Luminescent Proteins / Q000378:metabolism; D018129:Phosphatidylinositol Phosphates / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D011725:Pyridines / Q000494:pharmacology; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012697:Serine Endopeptidases / Q000737:chemistry / Q000378:metabolism; D014168:Transferrin / Q000378:metabolism; D033921:Vesicular Transport Proteins / Q000378:metabolism; D048051:p38 Mitogen-Activated Protein Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D007093:Imidazoles; D047908:Intracellular Signaling Peptides and Proteins; D008164:Luminescent Proteins; D018129:Phosphatidylinositol Phosphates; D011725:Pyridines; D011993:Recombinant Fusion Proteins; D014168:Transferrin; D033921:Vesicular Transport Proteins; C093846:early endosome antigen 1; C055525:phosphatidylinositol 3-phosphate; C054562:yellow fluorescent protein, Bacteria; D002772:Cholera Toxin; D006861:Hydrogen Peroxide; D058546:Class III Phosphatidylinositol 3-Kinases; D066246:ErbB Receptors; D048051:p38 Mitogen-Activated Protein Kinases; C116420:ZFYVE16 protein, human; D012697:Serine Endopeptidases; C093642:SB 203580