COPII-coated membranes function as transport carriers of intracellular procollagen I
Open Access
- 20 April 2017
- journal article
- video audio-media
- Published by Rockefeller University Press in The Journal of cell biology
- Vol. 216 (6), 1745-1759
- https://doi.org/10.1083/jcb.201702135
Abstract
The coat protein complex II (COPII) is essential for the transport of large cargo, such as 300-nm procollagen I (PC1) molecules, from the endoplasmic reticulum (ER) to the Golgi. Previous work has shown that the CUL3-KLHL12 complex increases the size of COPII vesicles at ER exit sites to more than 300 nm in diameter and accelerates the secretion of PC1. However, the role of large COPII vesicles as PC1 transport carriers was not unambiguously demonstrated. In this study, using stochastic optical reconstruction microscopy, correlated light electron microscopy, and live-cell imaging, we demonstrate the existence of mobile COPII-coated vesicles that completely encapsulate the cargo PC1 and are physically separated from ER. We also developed a cell-free COPII vesicle budding reaction that reconstitutes the capture of PC1 into large COPII vesicles. This process requires COPII proteins and the GTPase activity of the COPII subunit SAR1. We conclude that large COPII vesicles are bona fide carriers of PC1.Keywords
Funding Information
- Gordon and Betty Moore Foundation
- National Institutes of Health (1S10RR026866-01, 1S10OD018136-01)
- Howard Hughes Medical Institute
- University of California Berkeley
- Miller Institute for Basic Research in Science
- Lawrence Berkeley National Laboratory
- National Science Foundation (CHE-1554717)
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