Segregation of COPI-rich and anterograde-cargo-rich domains in endoplasmic-reticulum-to-Golgi transport complexes.
Membrane traffic between the endoplasmic reticulum (ER) and the Golgi complex is regulated by two vesicular coat complexes, COPII and COPI. COPII has been implicated in the selective packaging of anterograde cargo into coated transport vesicles budding from the ER [1]. In mammalian cells, these vesicles coalesce to form tubulo-vesicular transport complexes (TCs), which shuttle anterograde cargo from the ER to the Golgi complex [2] [3] [4]. In contrast, COPI-coated vesicles are proposed to mediate recycling of proteins from the Golgi complex to the ER [1] [5] [6] [7]. The binding of COPI to COPII-coated TCs [3] [8] [9], however, has led to the proposal that COPI binds to TCs and specifically packages recycling proteins into retrograde vesicles for return to the ER [3] [9]. To test this hypothesis, we tracked fluorescently tagged COPI and anterograde-transport markers simultaneously in living cells. COPI predominated on TCs shuttling anterograde cargo to the Golgi complex and was rarely observed on structures moving in directions consistent with retrograde transport. Furthermore, a progressive segregation of COPI-rich domains and anterograde-cargo-rich domains was observed in the TCs. This segregation and the directed motility of COPI-containing TCs were inhibited by antibodies that blocked COPI function. These observations, which are consistent with previous biochemical data [2] [9], suggest a role for COPI within TCs en route to the Golgi complex. By sequestering retrograde cargo in the anterograde-directed TCs, COPI couples the sorting of ER recycling proteins [10] to the transport of anterograde cargo.
Shima
D T
DT
Cell Biology Laboratory, Imperial Cancer Research Fund, 44, Lincoln's Inn Fields, London, WC2A 3PX, UK.
Scales
S J
SJ
Kreis
T E
TE
Pepperkok
R
R
D000818:Animals; D000911:Antibodies, Monoclonal / Q000494:pharmacology; D001693:Biological Transport, Active; D002232:Carbocyanines; D002522:Chlorocebus aethiops; D020755:Coat Protein Complex I / Q000037:antagonists & inhibitors / Q000276:immunology / Q000378:metabolism*; D004721:Endoplasmic Reticulum / Q000378:metabolism*; D005456:Fluorescent Dyes; D006056:Golgi Apparatus / Q000378:metabolism*; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins / Q000378:metabolism; D018613:Microscopy, Confocal; D008954:Models, Biological; D014709:Vero Cells
D000911:Antibodies, Monoclonal; D002232:Carbocyanines; D020755:Coat Protein Complex I; D005456:Fluorescent Dyes; D008164:Luminescent Proteins; C089187:cyanine dye 3; D049452:Green Fluorescent Proteins