1-Butanol targets the Golgi apparatus in tobacco BY-2 cells, but in a different way to Brefeldin A.
The effects of 1-butanol on the organelles of the early secretory pathway in tobacco BY-2 cells have been examined, because this primary alcohol is known to interfere with phospholipase D an enzyme whose activity contributes to COPI-vesicle formation. Since the fungal lactone Brefeldin A (BFA) also prevents COPI-vesicle production by the Golgi apparatus, the sequential and simultaneous application of these two inhibitors was also investigated. 1-Butanol, but not 2-butanol caused rapid changes in the morphology of the BY-2 Golgi apparatus resulting in extended curved cisternae. By contrast with BFA-treated cells, ER cisternae did not attach laterally to these structures, and ER-Golgi fusion hybrids were not obtained with 1-butanol. However, immunofluorescence microscopy revealed that 1-butanol, like BFA, elicited the release of the GTPase ARF1 from Golgi membranes. Washing out the butanol resulted in re-attachment of ARF1 and a recovery of Golgi stack morphology. BY-2 cells treated sequentially with 1-butanol then BFA (each 30 min), did not reveal any BFA-typical changes in Golgi structure. Cells treated first with BFA, then 1-butanol retained the typical ER-Golgi sandwich morphology induced by BFA, but were larger. When 1-butanol and BFA were added together (for a 30 min period), even larger Golgi aggregates were formed with, again, no ER attachments. Thus, although both inhibitors had the Golgi apparatus as their principle cytological target and both interfere with coatomer attachment, they differ in their ability to induce an interaction with the ER.
Langhans
Markus
M
Department of Cell Biology, Heidelberg Institute for Plant Sciences, University of Heidelberg, Im Neuenheimer Feld 230, D-69120 Heidelberg, Germany.
Robinson
David G
DG
D020001:1-Butanol / Q000494:pharmacology*; D020126:Brefeldin A / Q000494:pharmacology*; D002460:Cell Line; D006056:Golgi Apparatus / Q000187:drug effects*; D014026:Nicotiana / Q000166:cytology / Q000187:drug effects*
D020126:Brefeldin A; D020001:1-Butanol