Immuno-electron tomography of ER exit sites reveals the existence of free COPII-coated transport carriers

Nature Cell Biology
Dagmar ZeuschnerJ Klumperman

Abstract

Transport from the endoplasmic reticulum (ER) to the Golgi complex requires assembly of the COPII coat complex at ER exit sites. Recent studies have raised the question as to whether in mammalian cells COPII coats give rise to COPII-coated transport vesicles or instead form ER sub-domains that collect proteins for transport via non-coated carriers. To establish whether COPII-coated vesicles do exist in vivo, we developed approaches to combine quantitative immunogold labelling (to identify COPII) and three-dimensional electron tomography (to reconstruct entire membrane structures). In tomograms of both chemically fixed and high-pressure-frozen HepG2 cells, immuno-labelled COPII was found on ER-associated buds as well as on free approximately 50-nm diameter vesicles. In addition, we identified a novel type of COPII-coated structure that consists of partially COPII-coated, 150-200-nm long, dumb-bell-shaped tubules. Both COPII-coated carriers also contain the SNARE protein Sec22b, which is necessary for downstream fusion events. Our studies unambiguously establish the existence of free, bona fide COPII-coated transport carriers at the ER-Golgi interface, suggesting that assembly of COPII coats in vivo can result in vesicle formation.

References

Aug 1, 1991·The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society·A de GraafA J Verkleij
Jun 1, 1983·The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society·M C Willingham
Jan 1, 1996·Journal of Structural Biology·J R KremerJ R McIntosh
Oct 1, 1996·The Journal of Cell Biology·S I BannykhW E Balch
Jan 27, 1998·Journal of Structural Biology·A J KosterW Baumeister
Jan 27, 1998·Journal of Structural Biology·D N Mastronarde
Mar 24, 1999·The Journal of Cell Biology·M S LadinskyL A Staehelin
Jun 30, 2000·Current Opinion in Cell Biology·J Klumperman
Dec 10, 2003·Proceedings of the National Academy of Sciences of the United States of America·Gerbrand KosterMarileen Dogterom
Jan 28, 2004·Cell·Juan S Bonifacino, Benjamin S Glick
Oct 7, 2004·Current Opinion in Neurobiology·Richard D Leapman
Dec 8, 2004·Nature Cell Biology·Peter WatsonDavid J Stephens
Dec 22, 2004·The Journal of Cell Biology·Dalu Xu, Jesse C Hay
Jan 25, 2005·Nature Structural & Molecular Biology·Ken Sato, Akihiko Nakano
Jun 15, 2005·Annual Review of Biochemistry·Vladan LucićWolfgang Baumeister
Jun 25, 2005·Current Opinion in Cell Biology·J Christopher Fromme, Randy Schekman
Nov 30, 2005·Molecular Biology of the Cell·Jeremy C SimpsonRainer Pepperkok

❮ Previous
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Citations

Jul 24, 2008·Histochemistry and Cell Biology·Guillermo Vivero-SalmerónJosé A Martínez-Menárguez
Aug 4, 2009·Histochemistry and Cell Biology·Jason PiersonPeter J Peters
Jul 3, 2013·Histochemistry and Cell Biology·Emma Martínez-AlonsoJosé A Martínez-Menárguez
Jul 25, 2013·Histochemistry and Cell Biology·Kasey J DayBenjamin S Glick
Aug 21, 2013·Histochemistry and Cell Biology·Hong-Mei HanMarkus Grabenbauer
Apr 25, 2012·Cell and Tissue Research·Kristian PrydzJaakko Saraste
Dec 24, 2011·Nature Cell Biology·Giulia ZanettiRandy Schekman
Oct 20, 2007·Nature Protocols·Jan W Slot, Hans J Geuze
Sep 23, 2006·Nature Reviews. Molecular Cell Biology·Cemal GürkanWilliam E Balch
May 24, 2013·Nature Reviews. Molecular Cell Biology·Federica Brandizzi, Charles Barlowe
Jan 22, 2010·Molecular Biology of the Cell·Marvin BentleyJesse C Hay
Jun 28, 2011·Journal of Experimental Botany·Alexander LerichDavid G Robinson
Apr 20, 2011·Cold Spring Harbor Perspectives in Biology·Judith Klumperman
Jul 30, 2009·Journal of Cell Science·Helen HughesDavid J Stephens
Aug 6, 2009·The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society·Katia CorteseCarlo Tacchetti
Dec 23, 2006·Proceedings of the National Academy of Sciences of the United States of America·Bryon S DonohoeL Andrew Staehelin
Oct 1, 2013·Trends in Cell Biology·Rossella VendittiMaria Antonietta De Matteis
Dec 15, 2010·Molecular Membrane Biology·Katy Schmidt, David J Stephens
Oct 24, 2009·FEBS Letters·Annika Budnik, David J Stephens
Nov 3, 2009·FEBS Letters·Effrosyni Papanikou, Benjamin S Glick
Aug 1, 2007·The Journal of Cell Biology·Jian LiVictor W Hsu
Jun 16, 2010·The Journal of Cell Biology·Hesso FarhanHans-Peter Hauri
Aug 28, 2015·Proceedings of the National Academy of Sciences of the United States of America·Benjamin D EngelWolfgang Baumeister
Jul 26, 2011·Molecular and Cellular Neurosciences·José Ignacio ValenzuelaAndrés Couve
Mar 7, 2009·Annals of Anatomy = Anatomischer Anzeiger : Official Organ of the Anatomische Gesellschaft·Wiebke Möbius
Aug 10, 2007·Current Opinion in Cell Biology·R Luke WisemanWilliam E Balch
Aug 11, 2007·Seminars in Cell & Developmental Biology·Els Wessels, Jeremy C Simpson
Aug 19, 2007·Seminars in Cell & Developmental Biology·Semra J Kirk, Theresa H Ward
Mar 31, 2007·Current Opinion in Structural Biology·Scott M StaggWilliam E Balch
Feb 23, 2012·Scientific Reports·Kirsten BaciaRandy Schekman
Sep 17, 2009·Journal of Microscopy·A A Mironov, G V Beznoussenko
Aug 18, 2009·Molecular Microbiology·Reinhard Rachel
May 1, 2008·Journal of Microscopy·C Hagen, K Grünewald
Apr 25, 2007·Traffic·Elly van DonselaarJan W Slot
Jun 3, 2008·Traffic·Chris HawesImogen Sparkes
Jun 6, 2007·Traffic·Vytaute Starkuviene, Rainer Pepperkok

❮ Previous
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