Cholesterol Regulates Syntaxin 6 Trafficking at trans-Golgi Network Endosomal Boundaries
Meritxell Reverter,
Carles Rentero,
Ana Garcia-Melero,
Monira Hoque,
Sandra Vilà de Muga,
Anna Álvarez-Guaita,
James R.W. Conway,
Peta Wood,
Rose Cairns,
Lilia Lykopoulou,
Daniel Grinberg,
Lluïsa Vilageliu,
Marta Bosch,
Joerg Heeren,
Juan Blasi,
Paul Timpson,
Albert Pol,
Francesc Tebar,
Rachael Z. Murray,
Thomas Grewal,
Carlos Enrich
Affiliations
Meritxell Reverter
Departament de Biologia Cel·lular, Immunologia i Neurociències, Facultat de Medicina, Universitat de Barcelona, 08036 Barcelona, Spain
Carles Rentero
Departament de Biologia Cel·lular, Immunologia i Neurociències, Facultat de Medicina, Universitat de Barcelona, 08036 Barcelona, Spain
Ana Garcia-Melero
Departament de Biologia Cel·lular, Immunologia i Neurociències, Facultat de Medicina, Universitat de Barcelona, 08036 Barcelona, Spain
Monira Hoque
Faculty of Pharmacy, University of Sydney, Sydney, NSW 2006, Australia
Sandra Vilà de Muga
Departament de Biologia Cel·lular, Immunologia i Neurociències, Facultat de Medicina, Universitat de Barcelona, 08036 Barcelona, Spain
Anna Álvarez-Guaita
Departament de Biologia Cel·lular, Immunologia i Neurociències, Facultat de Medicina, Universitat de Barcelona, 08036 Barcelona, Spain
James R.W. Conway
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
Peta Wood
Faculty of Pharmacy, University of Sydney, Sydney, NSW 2006, Australia
Rose Cairns
Faculty of Pharmacy, University of Sydney, Sydney, NSW 2006, Australia
Lilia Lykopoulou
First Department of Pediatrics, University of Athens, Aghia Sofia Children’s Hospital, 11527 Athens, Greece
Daniel Grinberg
Departament de Genètica, Facultat de Biologia, Universitat de Barcelona, CIBERER, IBUB, 08028 Barcelona, Spain
Lluïsa Vilageliu
Departament de Genètica, Facultat de Biologia, Universitat de Barcelona, CIBERER, IBUB, 08028 Barcelona, Spain
Marta Bosch
Centre de Recerca Biomèdica CELLEX, Institut d’Investigacions Biomèdiques August Pi i Sunyer (IDIBAPS), 08036 Barcelona, Spain
Joerg Heeren
Department of Biochemistry and Molecular Biology II. Molecular Cell Biology, University Medical Center Hamburg-Eppendorf, 20246 Hamburg, Germany
Juan Blasi
Department of Pathology and Experimental Therapeutics, IDIBELL-University of Barcelona, L’Hospitalet de Llobregat, 08907 Barcelona, Spain
Paul Timpson
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
Albert Pol
Departament de Biologia Cel·lular, Immunologia i Neurociències, Facultat de Medicina, Universitat de Barcelona, 08036 Barcelona, Spain
Francesc Tebar
Departament de Biologia Cel·lular, Immunologia i Neurociències, Facultat de Medicina, Universitat de Barcelona, 08036 Barcelona, Spain
Rachael Z. Murray
Tissue Repair and Regeneration Program, Institute of Health and Biomedical, Innovation, Queensland University of Technology, Brisbane, QLD 4095, Australia
Thomas Grewal
Faculty of Pharmacy, University of Sydney, Sydney, NSW 2006, Australia
Carlos Enrich
Departament de Biologia Cel·lular, Immunologia i Neurociències, Facultat de Medicina, Universitat de Barcelona, 08036 Barcelona, Spain
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.
