title: Cholesterol regulates Syntaxin 6 trafficking at trans-Golgi network endosomal boundaries.
abstract: Inhibition of cholesterol export from late endosomes causes cellular cholesterol imbalance, including cholesterol depletion in the trans-Golgi network (TGN). Here, using Chinese hamster ovary (CHO) Niemann-Pick type C1 (NPC1) mutant cell lines and human NPC1 mutant fibroblasts, we show that altered cholesterol levels at the TGN/endosome boundaries trigger Syntaxin 6 (Stx6) accumulation into VAMP3, transferrin, and Rab11-positive recycling endosomes (REs). This increases Stx6/VAMP3 interaction and interferes with the recycling of αVβ3 and α5β1 integrins and cell migration, possibly in a Stx6-dependent manner. In NPC1 mutant cells, restoration of cholesterol levels in the TGN, but not inhibition of VAMP3, restores the steady-state localization of Stx6 in the TGN. Furthermore, elevation of RE cholesterol is associated with increased amounts of Stx6 in RE. Hence, the fine-tuning of cholesterol levels at the TGN-RE boundaries together with a subset of cholesterol-sensitive SNARE proteins may play a regulatory role in cell migration and invasion.
authors.affiliation: Departament de Biologia Cel·lular, Immunologia i Neurociències, Facultat de Medicina, Universitat de Barcelona, 08036 Barcelona, Spain.
authors.forename: Meritxell
authors.initials: M
authors.lastname: Reverter
authors.affiliation: Departament de Biologia Cel·lular, Immunologia i Neurociències, Facultat de Medicina, Universitat de Barcelona, 08036 Barcelona, Spain.
authors.forename: Carles
authors.initials: C
authors.lastname: Rentero
authors.affiliation: Departament de Biologia Cel·lular, Immunologia i Neurociències, Facultat de Medicina, Universitat de Barcelona, 08036 Barcelona, Spain.
authors.forename: Ana
authors.initials: A
authors.lastname: Garcia-Melero
authors.affiliation: Faculty of Pharmacy, University of Sydney, Sydney, NSW 2006, Australia.
authors.forename: Monira
authors.initials: M
authors.lastname: Hoque
authors.affiliation: Departament de Biologia Cel·lular, Immunologia i Neurociències, Facultat de Medicina, Universitat de Barcelona, 08036 Barcelona, Spain.
authors.forename: Sandra
authors.initials: S
authors.lastname: Vilà de Muga
authors.affiliation: Departament de Biologia Cel·lular, Immunologia i Neurociències, Facultat de Medicina, Universitat de Barcelona, 08036 Barcelona, Spain.
authors.forename: Anna
authors.initials: A
authors.lastname: Alvarez-Guaita
authors.affiliation: Garvan Institute of Medical Research and Kinghorn Cancer Centre, Cancer Research Program, St. Vincent's Clinical School, Faculty of Medicine, University of New South Wales, Sydney, NSW 2010, Australia.
authors.forename: James R W
authors.initials: JR
authors.lastname: Conway
authors.affiliation: Faculty of Pharmacy, University of Sydney, Sydney, NSW 2006, Australia.
authors.forename: Peta
authors.initials: P
authors.lastname: Wood
authors.affiliation: Faculty of Pharmacy, University of Sydney, Sydney, NSW 2006, Australia.
authors.forename: Rose
authors.initials: R
authors.lastname: Cairns
authors.affiliation: First Department of Pediatrics, University of Athens, Aghia Sofia Children's Hospital, 11527 Athens, Greece.
authors.forename: Lilia
authors.initials: L
authors.lastname: Lykopoulou
authors.affiliation: Departament de Genètica, Facultat de Biologia, Universitat de Barcelona, CIBERER, IBUB, 08028 Barcelona, Spain.
authors.forename: Daniel
authors.initials: D
authors.lastname: Grinberg
authors.affiliation: Departament de Genètica, Facultat de Biologia, Universitat de Barcelona, CIBERER, IBUB, 08028 Barcelona, Spain.
authors.forename: Lluïsa
authors.initials: L
authors.lastname: Vilageliu
authors.affiliation: Centre de Recerca Biomèdica CELLEX, Institut d'Investigacions Biomèdiques August Pi i Sunyer (IDIBAPS), 08036 Barcelona, Spain.
authors.forename: Marta
authors.initials: M
authors.lastname: Bosch
authors.affiliation: Department of Biochemistry and Molecular Biology II. Molecular Cell Biology, University Medical Center Hamburg-Eppendorf, 20246 Hamburg, Germany.
authors.forename: Joerg
authors.initials: J
authors.lastname: Heeren
authors.affiliation: Department of Pathology and Experimental Therapeutics, IDIBELL-University of Barcelona, L'Hospitalet de Llobregat, 08907 Barcelona, Spain.
authors.forename: Juan
authors.initials: J
authors.lastname: Blasi
authors.affiliation: Garvan Institute of Medical Research and Kinghorn Cancer Centre, Cancer Research Program, St. Vincent's Clinical School, Faculty of Medicine, University of New South Wales, Sydney, NSW 2010, Australia.
authors.forename: Paul
authors.initials: P
authors.lastname: Timpson
authors.affiliation: Departament de Biologia Cel·lular, Immunologia i Neurociències, Facultat de Medicina, Universitat de Barcelona, 08036 Barcelona, Spain; Centre de Recerca Biomèdica CELLEX, Institut d'Investigacions Biomèdiques August Pi i Sunyer (IDIBAPS), 08036 Barcelona, Spain; Institució Catalana de Recerca i Estudis Avaçats (ICREA), 08010 Barcelona, Spain.
authors.forename: Albert
authors.initials: A
authors.lastname: Pol
authors.affiliation: Departament de Biologia Cel·lular, Immunologia i Neurociències, Facultat de Medicina, Universitat de Barcelona, 08036 Barcelona, Spain; Centre de Recerca Biomèdica CELLEX, Institut d'Investigacions Biomèdiques August Pi i Sunyer (IDIBAPS), 08036 Barcelona, Spain.
authors.forename: Francesc
authors.initials: F
authors.lastname: Tebar
authors.affiliation: Tissue Repair and Regeneration Program, Institute of Health and Biomedical, Innovation, Queensland University of Technology, Brisbane, QLD 4095, Australia.
authors.forename: Rachael Z
authors.initials: RZ
authors.lastname: Murray
authors.affiliation: Faculty of Pharmacy, University of Sydney, Sydney, NSW 2006, Australia. Electronic address: thomas.grewal@sydney.edu.au.
authors.forename: Thomas
authors.initials: T
authors.lastname: Grewal
authors.affiliation: Departament de Biologia Cel·lular, Immunologia i Neurociències, Facultat de Medicina, Universitat de Barcelona, 08036 Barcelona, Spain; Centre de Recerca Biomèdica CELLEX, Institut d'Investigacions Biomèdiques August Pi i Sunyer (IDIBAPS), 08036 Barcelona, Spain. Electronic address: enrich@ub.edu.
authors.forename: Carlos
authors.initials: C
authors.lastname: Enrich
mesh_terms: D000818:Animals; D016466:CHO Cells; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism; D002465:Cell Movement; D002784:Cholesterol / Q000378:metabolism*; D006224:Cricetinae; D003412:Cricetulus; D011992:Endosomes / Q000378:metabolism*; D006801:Humans; D039081:Integrin alpha5beta1 / Q000378:metabolism; D047908:Intracellular Signaling Peptides and Proteins; D008562:Membrane Glycoproteins / Q000235:genetics / Q000378:metabolism; D000077296:Niemann-Pick C1 Protein; D011485:Protein Binding; D021381:Protein Transport; D050765:Qa-SNARE Proteins / Q000737:chemistry / Q000378:metabolism*; D019030:Receptors, Vitronectin / Q000378:metabolism; D050600:SNARE Proteins / Q000378:metabolism; D051040:Vesicle-Associated Membrane Protein 3 / Q000737:chemistry / Q000378:metabolism; D020691:rab GTP-Binding Proteins / Q000378:metabolism; D021601:trans-Golgi Network / Q000378:metabolism*; D000099193:rab11 GTP-Binding Proteins
chemical_list: D002352:Carrier Proteins; D002784:Cholesterol; D039081:Integrin alpha5beta1; D047908:Intracellular Signaling Peptides and Proteins; D008562:Membrane Glycoproteins; D000077296:Niemann-Pick C1 Protein; D050765:Qa-SNARE Proteins; D019030:Receptors, Vitronectin; D050600:SNARE Proteins; D051040:Vesicle-Associated Membrane Protein 3; D020691:rab GTP-Binding Proteins; D000099193:rab11 GTP-Binding Proteins; C106881:NPC1 protein, human; C094340:integrin alphavbeta1