Menu

Close

ER-to-Golgi trafficking of procollagen in the absence of large carriers.

Janine McCaughey, Nicola Stevenson, Stephen Cross, David Stephens

Preprint posted on June 05, 2018 https://www.biorxiv.org/content/early/2018/06/05/339804

Article now published in The Journal of Cell Biology at http://dx.doi.org/10.1083/jcb.201806035

Light at the end of the tunnel? A new reporter for pulsed collagen secretion provides support for a direct route from ER to Golgi

Selected by Gautam Dey

Context

Multicellularity brings with it myriad advantages but also a problem of scales- millimetre-scale tissue architecture must be wrought by cells themselves often only tens of microns across. Nowhere is this more evident than at one of the early steps in the creation of the extra-cellular matrix (ECM). Collagen, typically the major ECM component (and the most abundant protein in the human body), is folded and processed in the ER and subsequently the Golgi before transport to the cell surface1.

The first steps in collagen secretion at ER exit sites are orchestrated by a protein called TANGO12 and its interactors, with the participation of the canonical ER-Golgi carrier coat protein complex COPII. One major wrinkle in the story, though: ER-localised procollagen takes the form of a gigantic, rigid rod 300 nm long. How on earth do procollagens fit into COPII-coated vesicles less than 100 nm across? Competing models for how this might reasonably occur- giant vesicular carriers3 or direct tunnels/channels1,4,5 between the ERGIC and Golgi- have both received some experimental support.

 

Key advance

The authors designed a novel live-cell reporter for human COL1A1, placing an N-terminal monomeric GFP tag upstream of the pro-peptide cleavage site (to eliminate signals from extracellular collagen) in combination with a Streptavidin Binding Peptide (SBP, to utilise in combination with an ER-resident streptavidin “hook”6) and selecting for stable, low expression. The critical assay involved monitoring synchronised pulses of collagen transport by confocal imaging from ER to Golgi by treating cells with ascorbate (to assist in collagen folding) and biotin (to release the COL1A1 from the hook).

The authors were able to track emptying of the ER and filling of the Golgi on a 20-30-minute timescale with high time and spatial resolution but were unable to detect any large vesicular carriers. Any large (or small) COL1A1 labelled structures not classified as bona-fide Golgi colocalised only with ER markers.

Figure 1: Panels reproduced from Figure 2D of McCaughey et al., 2018 under a CC-BY-NC-ND 4.0 international license, representing the collagen reporter (GFP-COL) and a trans-Golgi marker (ST-Cherry) during a pulse-chase assay.

 

 

Why I chose this preprint  

Recent advances in light microscopy and labelling techniques7 will have a broad impact on the way we study the cell biology of cargo flux between dynamic eukaryotic membrane compartments. Super-resolution imaging (albeit largely in fixed cells) has already made its mark on the collagen secretion field3,8. This preprint seems like another step in the right direction!

 

Questions for the future?

I was fascinated by the way the filling of the Golgi with collagen (as in Figure 2D excerpted above and in many other examples in the preprint) appears to actually physically reshape the Golgi stacks over time. Could the authors’ assay double up as a way to investigate the role of cargo in determining compartment structure, and could such experiments be aided by a more quantitative approach to tracking the dynamic evolution of compartments in space (e.g. machine learning)?

 

References:

  1. Malhotra, V. & Erlmann, P. The Pathway of Collagen Secretion. Annu. Rev. Cell Dev. Biol. 31, 109–124 (2015).
  2. Saito, K. et al. TANGO1 Facilitates Cargo Loading at Endoplasmic Reticulum Exit Sites. Cell 136, 891–902 (2009).
  3. Gorur, A. et al. COPII-coated membranes function as transport carriers of intracellular procollagen I. J. Cell Biol. 216, 1745–1759 (2017).
  4. Beznoussenko, G. V et al. Transport of soluble proteins through the Golgi occurs by diffusion via continuities across cisternae. Elife 3, (2014).
  5. Kurokawa, K., Okamoto, M. & Nakano, A. Contact of cis-Golgi with ER exit sites executes cargo capture and delivery from the ER. Nat. Commun. 5, 3653 (2014).
  6. Boncompain, G. et al. Synchronization of secretory protein traffic in populations of cells. Nat. Methods 9, 493–498 (2012).
  7. Pickard, A. et al. Collagen assembly and turnover imaged with a CRISPR-Cas9 engineered Dendra2 tag. bioRxiv 331496 (2018). doi:10.1101/331496
  8. Raote, I. et al. TANGO1 assembles into rings around COPII coats at ER exit sites. J. Cell Biol. 216, 901–909 (2017).

Tags: er-golgi transport, membrane trafficking

Posted on: 26th July 2018 , updated on: 30th July 2018

Read preprint (No Ratings Yet)




  • Author's response

    Janine McCaughey, Nicola Stevenson, and David Stephens shared

    We appreciate Gautam highlighting our work. We do think that the new biology that we define in this paper will be of broad interest. Our new controllable trafficking tool provides an opportunity to define many different aspects of procollagen trafficking. The observation that the trafficking of our mGFP-procollagen through the Golgi might “physically reshape the Golgi stacks over time” is very intriguing. The Golgi is highly dynamic and so we are not drawing that conclusion at present. This would of course require extensive analysis by both light and electron microscopy and we are hoping to work towards that in the near future. Certainly, the possibility of applying machine learning to such image analysis methods is an exciting opportunity and we would love to talk to anyone working in that area.

    1 comment

    2 years

    David Stephens

    A revised version of our preprint is now available (17 Oct 2018).
    https://www.biorxiv.org/content/early/2018/10/17/339804

    Have your say

    Your email address will not be published. Required fields are marked *

    This site uses Akismet to reduce spam. Learn how your comment data is processed.

    Sign up to customise the site to your preferences and to receive alerts

    Register here

    preLists in the cell biology category:

    BioMalPar XVI: Biology and Pathology of the Malaria Parasite

    [under construction] Preprints presented at the (fully virtual) EMBL BioMalPar XVI, 17-18 May 2020 #emblmalaria

     



    List by Gautam Dey, Samantha Seah

    1

    Cell Polarity

    Recent research from the field of cell polarity is summarized in this list of preprints. It comprises of studies focusing on various forms of cell polarity ranging from epithelial polarity, planar cell polarity to front-to-rear polarity.

     



    List by Yamini Ravichandran

    TAGC 2020

    Preprints recently presented at the virtual Allied Genetics Conference, April 22-26, 2020. #TAGC20

     



    List by Maiko Kitaoka, Madhuja Samaddar, Miguel V. Almeida, Sejal Davla, Jennifer Ann Black, Gautam Dey

    3D Gastruloids

    A curated list of preprints related to Gastruloids (in vitro models of early development obtained by 3D aggregation of embryonic cells)

     



    List by Paul Gerald L. Sanchez and Stefano Vianello

    ECFG15 – Fungal biology

    Preprints presented at 15th European Conference on Fungal Genetics 17-20 February 2020 Rome

     



    List by Hiral Shah

    ASCB EMBO Annual Meeting 2019

    A collection of preprints presented at the 2019 ASCB EMBO Meeting in Washington, DC (December 7-11)

     



    List by Madhuja Samaddar, Ramona Jühlen, Amanda Haage, Laura McCormick, Maiko Kitaoka

    EMBL Seeing is Believing – Imaging the Molecular Processes of Life

    Preprints discussed at the 2019 edition of Seeing is Believing, at EMBL Heidelberg from the 9th-12th October 2019

     



    List by Gautam Dey

    Autophagy

    Preprints on autophagy and lysosomal degradation and its role in neurodegeneration and disease. Includes molecular mechanisms, upstream signalling and regulation as well as studies on pharmaceutical interventions to upregulate the process.

     



    List by Sandra Malmgren Hill

    Lung Disease and Regeneration

    This preprint list compiles highlights from the field of lung biology.

     



    List by Rob Hynds

    Cellular metabolism

    A curated list of preprints related to cellular metabolism at Biorxiv by Pablo Ranea Robles from the Prelights community. Special interest on lipid metabolism, peroxisomes and mitochondria.

     



    List by Pablo Ranea Robles

    BSCB/BSDB Annual Meeting 2019

    Preprints presented at the BSCB/BSDB Annual Meeting 2019

     



    List by Gautam Dey

    MitoList

    This list of preprints is focused on work expanding our knowledge on mitochondria in any organism, tissue or cell type, from the normal biology to the pathology.

     



    List by Sandra Franco Iborra

    Biophysical Society Annual Meeting 2019

    Few of the preprints that were discussed in the recent BPS annual meeting at Baltimore, USA

     



    List by Joseph Jose Thottacherry

    ASCB/EMBO Annual Meeting 2018

    This list relates to preprints that were discussed at the recent ASCB conference.

     



    List by Gautam Dey, Amanda Haage

    Also in the developmental biology category:

    Cell Polarity

    Recent research from the field of cell polarity is summarized in this list of preprints. It comprises of studies focusing on various forms of cell polarity ranging from epithelial polarity, planar cell polarity to front-to-rear polarity.

     



    List by Yamini Ravichandran

    TAGC 2020

    Preprints recently presented at the virtual Allied Genetics Conference, April 22-26, 2020. #TAGC20

     



    List by Maiko Kitaoka, Madhuja Samaddar, Miguel V. Almeida, Sejal Davla, Jennifer Ann Black, Gautam Dey

    3D Gastruloids

    A curated list of preprints related to Gastruloids (in vitro models of early development obtained by 3D aggregation of embryonic cells)

     



    List by Paul Gerald L. Sanchez and Stefano Vianello

    ASCB EMBO Annual Meeting 2019

    A collection of preprints presented at the 2019 ASCB EMBO Meeting in Washington, DC (December 7-11)

     



    List by Madhuja Samaddar, Ramona Jühlen, Amanda Haage, Laura McCormick, Maiko Kitaoka

    EDBC Alicante 2019

    Preprints presented at the European Developmental Biology Congress (EDBC) in Alicante, October 23-26 2019.

     



    List by Sergio Menchero, Jesus Victorino, Teresa Rayon, Irepan Salvador-Martinez

    EMBL Seeing is Believing – Imaging the Molecular Processes of Life

    Preprints discussed at the 2019 edition of Seeing is Believing, at EMBL Heidelberg from the 9th-12th October 2019

     



    List by Gautam Dey

    SDB 78th Annual Meeting 2019

    A curation of the preprints presented at the SDB meeting in Boston, July 26-30 2019. The preList will be updated throughout the duration of the meeting.

     



    List by Alex Eve

    Lung Disease and Regeneration

    This preprint list compiles highlights from the field of lung biology.

     



    List by Rob Hynds

    Young Embryologist Network Conference 2019

    Preprints presented at the Young Embryologist Network 2019 conference, 13 May, The Francis Crick Institute, London

     



    List by Alex Eve

    Pattern formation during development

    The aim of this preList is to integrate results about the mechanisms that govern patterning during development, from genes implicated in the processes to theoritical models of pattern formation in nature.

     



    List by Alexa Sadier

    BSCB/BSDB Annual Meeting 2019

    Preprints presented at the BSCB/BSDB Annual Meeting 2019

     



    List by Gautam Dey

    Zebrafish immunology

    A compilation of cutting-edge research that uses the zebrafish as a model system to elucidate novel immunological mechanisms in health and disease.

     



    List by Shikha Nayar
    Close