Dissecting Functions of the Conserved Oligomeric Golgi Tethering Complex Using a Cell-Free Assay
Version of Record online: 31 OCT 2013
©2013 The Authors. Traffic published by John Wiley & Sons Ltd.
This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
Volume 15, Issue 1, pages 12–21, January 2014
How to Cite
Cottam, N. P., Wilson, K. M., Ng, B. G., Körner, C., Freeze, H. H. and Ungar, D. (2014), Dissecting Functions of the Conserved Oligomeric Golgi Tethering Complex Using a Cell-Free Assay. Traffic, 15: 12–21. doi: 10.1111/tra.12128
- Issue online: 6 DEC 2013
- Version of Record online: 31 OCT 2013
- Accepted manuscript online: 4 OCT 2013 09:56AM EST
- Manuscript Accepted: 4 OCT 2013
- Manuscript Revised: 2 OCT 2013
- Manuscript Received: 5 MAY 2013
- BBSRC PhD Studentship
- Marie Curie Grant. Grant Number: 201098
- NIH. Grant Number: R01DK55615
- The Rocket Fund
- cell-free reconstitution;
- congenital disorders of glycosylation;
- conserved oligomeric Golgi complex;
- glycosylation enzyme sorting;
- Golgi apparatus;
- vesicle tethering
Vesicle transport sorts proteins between compartments and is thereby responsible for generating the non-uniform protein distribution along the eukaryotic secretory and endocytic pathways. The mechanistic details of specific vesicle targeting are not yet well characterized at the molecular level. We have developed a cell-free assay that reconstitutes vesicle targeting utilizing the recycling of resident enzymes within the Golgi apparatus. The assay has physiological properties, and could be used to show that the two lobes of the conserved oligomeric Golgi tethering complex play antagonistic roles in trans-Golgi vesicle targeting. Moreover, we can show that the assay is sensitive to several different congenital defects that disrupt Golgi function and therefore cause glycosylation disorders. Consequently, this assay will allow mechanistic insight into the targeting step of vesicle transport at the Golgi, and could also be useful for characterizing some novel cases of congenital glycosylation disorders.