IMMUNOCYTOCHEMICAL LOCALIZATION OF BETA-COP TO THE ER-GOLGI BOUNDARY AND THE TGN

Recent data strongly suggest that the coatomer (COP) complex is involv ed in membrane transport between the ER and Golgi complex. This vesicu lar coat has been implicated in ER to Golgi, in intra Golgi as well as in Golgi to ER traffic. In this study we present a detailed immunocyt ochemical analysis of the distribution of beta-COP in different tissue culture cells. Our results extend previous studies by showing, using electron microscopy, that beta-COP accumulates on vesicular profiles a nd buds in the intermediate compartment (IC) under conditions that blo ck ER to Golgi transport (15 degrees C), Importantly, under these cond itions beta-COP co-localizes on these structures with a passenger prot ein, the membrane glycoprotein of vesicular stomatis virus (ts-O45-G). Furthermore, quantitative immunofluorescence microscopy of cells with ts-045-G accumulated in the ER, IC and trans-Golgi network, shifted b riefly to the permissive temperature, showed that beta-COP was associa ted with many of the putative transport intermediates containing the v iral glycoprotein which is in transit between the ER/IC and the cis-Go lgi. The simplest interpretation of these data is that COP-coated vesi cles are involved in anterograde transport of ts-045-G from the IC to the Golgi complex. Since many putative COP vesicles lacked the G prote in following release of the 15 degrees C block this pool could be invo lved in retrograde transport. We also show that beta-COP is present on the membranes of the trans-Golgi network. However, in contrast to the ER-Golgi boundary, we could find no convincing evidence that this poo l of beta-COP is associated with buds or trans-Golgi network-derived t ransport vesicles.