Imaging of procollagen transport reveals COPI-dependent cargo sorting during ER-to-Golgi transport in mammalian cells.
We have examined the ER-to-Golgi transport of procollagen, which, when assembled in the lumen of the ER, is thought to be physically too large to fit in classically described 60-80 nm COPI- and COPII-coated transport vesicles. We found that procollagen exits the ER via COPII- coated ER exit sites and is transported to the Golgi along microtubules in defined transport complexes. These procollagen-containing transport complexes are, however, distinct from those containing other cargo proteins like ERGIC-53 and ts-045-G. Furthermore, they do not label for the COPI coat complex in contrast to those containing ts-045-G. Inhibition of COPII or COPI function before addition of ascorbate, which is required for the folding of procollagen, inhibits export of procollagen from the ER. Inactivation of COPI coat function after addition of ascorbate results in the localisation of procollagen to transport complexes that now also contain ERGIC-53 and are inhibited in their transport to the Golgi complex. These data reveal the existence of an early COPI-dependent, pre-Golgi cargo sorting step in mammalian cells.
Stephens
David J
DJ
Cell Biology and Cell Biophysics Programme, EMBL Heidelberg, Meyerhofstrasse 1, 69117 Heidelberg, Germany. david.stephens@bristol.ac.uk
Pepperkok
Rainer
R
D000818:Animals; D001693:Biological Transport, Active; D002478:Cells, Cultured; D002522:Chlorocebus aethiops; D020755:Coat Protein Complex I / Q000502:physiology*; D004721:Endoplasmic Reticulum / Q000378:metabolism*; D006056:Golgi Apparatus / Q000378:metabolism*; D049452:Green Fluorescent Proteins; D006367:HeLa Cells; D006801:Humans; D008164:Luminescent Proteins / Q000737:chemistry; D008322:Mammals; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D011347:Procollagen / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011487:Protein Conformation; D021381:Protein Transport; D022161:Transport Vesicles / Q000502:physiology; D014709:Vero Cells
D020755:Coat Protein Complex I; D008164:Luminescent Proteins; D011347:Procollagen; D049452:Green Fluorescent Proteins