Dynamic distribution of the Golgi marker thiamine pyrophosphatase is modulated by brefeldin A in rat hepatoma cells.
Cytochemical electron microscopy of cultured rat hepatoma cells (AH-130) demonstrated that thiamine pyrophosphatase (TPPase) activity was localized in the Golgi complex. When the cells were treated with brefeldin A (BFA, 2.5 micrograms/ml) for 10 min, the characteristic structure of the Golgi stack was no longer observed, and TPPase was cytochemically stained in the vesicular and tubular structures scattered in the cytoplasm. A longer exposure of the cells to the drug (20 min to 1 h) resulted in the distribution of the TPPase activity in the endoplasmic reticulum (ER) and nuclear envelope. Such an unusual distribution of the enzyme activity, however, was reversible even in the presence of BFA. At 2 h after the exposure, the TPPase activity disappeared from the ER and was concentrated again in the vesicular and tubular structures. The enzyme activity was finally localized in the Golgi complex which was reassembled by 4 h after the exposure. The reversible effect of BFA may be due to a possible metabolism of the drug into an inert form during the incubation. Taken together, these results indicate that BFA causes a rapid disassembly of the Golgi complex and redistribution of the marker enzyme TPPase into the ER including the nuclear envelope. The spontaneous reversibility of the drug effect also favors a dynamic recycling of the Golgi marker between the ER and the Golgi complex under the conditions used.
Fujiwara
T
T
Department of Biochemistry, Fukuoka University School of Medicine Jonanku, Japan.
Oda
K
K
Ikehara
Y
Y
D000818:Animals; D020126:Brefeldin A; D003517:Cyclopentanes / Q000494:pharmacology*; D006056:Golgi Apparatus / Q000187:drug effects / Q000201:enzymology* / Q000648:ultrastructure; D006651:Histocytochemistry; D008114:Liver Neoplasms, Experimental / Q000201:enzymology*; D008854:Microscopy, Electron; D011755:Pyrophosphatases / Q000378:metabolism*; D051381:Rats; D013834:Thiamine Pyrophosphatase / Q000378:metabolism*; D014407:Tumor Cells, Cultured
D003517:Cyclopentanes; D020126:Brefeldin A; D011755:Pyrophosphatases; D013834:Thiamine Pyrophosphatase