Fluorescence Lifetime Imaging Microscopy reveals rerouting of SNARE trafficking driving dendritic cell activation

ELife
Daniëlle Rianne José VerboogenGeert van den Bogaart

Abstract

SNARE proteins play a crucial role in intracellular trafficking by catalyzing membrane fusion, but assigning SNAREs to specific intracellular transport routes is challenging with current techniques. We developed a novel Förster resonance energy transfer-fluorescence lifetime imaging microscopy (FRET-FLIM)-based technique allowing visualization of real-time local interactions of fluorescently tagged SNARE proteins in live cells. We used FRET-FLIM to delineate the trafficking steps underlying the release of the inflammatory cytokine interleukin-6 (IL-6) from human blood-derived dendritic cells. We found that activation of dendritic cells by bacterial lipopolysaccharide leads to increased FRET of fluorescently labeled syntaxin 4 with VAMP3 specifically at the plasma membrane, indicating increased SNARE complex formation, whereas FRET with other tested SNAREs was unaltered. Our results revealed that SNARE complexing is a key regulatory step for cytokine production by immune cells and prove the applicability of FRET-FLIM for visualizing SNARE complexes in live cells with subcellular spatial resolution.

References

Dec 1, 1996·Molecular Biology of the Cell·S H LowK E Mostov
Apr 1, 1998·Nature·J Banchereau, R M Steinman
May 3, 2000·The Journal of Cell Biology·L BajnoS Grinstein
Apr 18, 2001·Trends in Cell Biology·H R Pelham
Jun 2, 2001·The Journal of Biological Chemistry·O GriesbeckR Y Tsien
May 30, 2003·The Biochemical Journal·Todd D SchrawSidney W Whiteheart
Nov 5, 2003·Nature Biotechnology·Elizabeth A Jares-Erijman, Thomas M Jovin
Feb 22, 2005·Current Opinion in Biotechnology·Horst Wallrabe, Ammasi Periasamy
May 17, 2005·Biochimica Et Biophysica Acta·Wanjin Hong
Oct 7, 2005·Molecular Biology of the Cell·Martin B A ter BeestKeith E Mostov
Nov 12, 2005·Science·Rachael Z MurrayJennifer L Stow
Aug 9, 2006·Biochemistry·Xiaokun ShuS James Remington
Aug 17, 2006·Nature Reviews. Molecular Cell Biology·Reinhard Jahn, Richard H Scheller
Feb 1, 2007·The Journal of Biological Chemistry·Colin RickmanRory R Duncan
Jun 8, 2007·The Journal of Biological Chemistry·Mari ValkonenRory R Duncan
Jul 4, 2007·The Journal of Cell Biology·Anthony P MandersonJennifer L Stow
Dec 7, 2007·Journal of Cell Science·Claire N MedineRory R Duncan
Feb 23, 2008·The Journal of Immunology : Official Journal of the American Association of Immunologists·Yong Hou Sunny HoSiew Heng Wong
Sep 5, 2008·Proceedings of the National Academy of Sciences of the United States of America·Buz BarstowSol M Gruner
Mar 10, 2009·Immunobiology·Jennifer L StowDaniele Sangermani
Jul 3, 2009·Nature·Alexander SteinReinhard Jahn
Jul 18, 2009·Biochemical and Biophysical Research Communications·Yong Hou Sunny HoSiew Heng Wong
Jan 23, 2010·The Journal of Biological Chemistry·Colin RickmanRory R Duncan
Feb 9, 2010·Nature Structural & Molecular Biology·Geert van den BogaartReinhard Jahn
Aug 12, 2010·Physiology·Amanda C Stanley, Paige Lacy
Oct 11, 2011·Molecular Immunology·Simon P C FrankAxel Lorentz
May 16, 2012·FEBS Letters·Mateusz Putyrski, Carsten Schultz
Jun 30, 2012·Nature Methods·Johannes SchindelinAlbert Cardona
Aug 30, 2012·Nature Methods·Caroline A SchneiderKevin W Eliceiri
Mar 29, 2013·The Journal of Neuroscience : the Official Journal of the Society for Neuroscience·Vadim DegtyarRobert S Zucker
Oct 23, 2013·Journal of Leukocyte Biology·Laura Gómez-JaramilloAntonio Campos-Caro
Nov 12, 2013·Current Topics in Membranes·Geert van den BogaartReinhard Jahn
Nov 12, 2014·Frontiers in Immunology·Rachael Zoe Murray, Jennifer Lea Stow

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Citations

Mar 24, 2018·FEBS Letters·Daniëlle R J VerboogenGeert van den Bogaart
May 24, 2018·Physiological Reviews·Ilse DingjanGeert van den Bogaart
Mar 17, 2018·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Omar A Z TutakhelJenny van der Wijst
Jul 18, 2018·Journal of Molecular Cell Biology·Daniëlle R J VerboogenGeert van den Bogaart
Oct 10, 2019·Traffic·Michael J Clague, Sylvie Urbé
Oct 12, 2019·Journal of Cell Science·Natalia H ReveloGeert van den Bogaart
Mar 9, 2021·Frontiers in Immunology·Maksim V BaranovGeert van den Bogaart
Dec 4, 2021·Immunological Reviews·Berislav BošnjakSwantje I Hammerschmidt

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Methods Mentioned

BETA
co-immunoprecipitation
FRET
confocal microscopy
Fluorescence
ELISA
Transfection

Software Mentioned

TRI2
Spechron
OriginPro2016
FIJI ImageJ
PT32ICS

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