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Vol. 10, Issue 12, 4021-4032, December 1999
Department of Molecular Biology and Genetics, Cornell University,
Ithaca, New York 14853
Evidence has been presented both for and against obligate
retrograde movement of resident Golgi proteins through the endoplasmic reticulum (ER) during nocodazole-induced Golgi ministack formation. Here, we studied the nocodazole-induced formation of ministacks using
phospholipase A2 (PLA2) antagonists, which have
been shown previously to inhibit brefeldin A-stimulated
Golgi-to-ER retrograde transport. Examination of clone 9 rat
hepatocytes by immunofluorescence and immunoelectron microscopy
revealed that a subset of PLA2 antagonists prevented
nocodazole-induced ministack formation by inhibiting two different
trafficking pathways for resident Golgi enzymes; at 25 µM, retrograde
Golgi-to-ER transport was inhibited, whereas at 5 µM, Golgi-to-ER
trafficking was permitted, but resident Golgi enzymes accumulated in
the ER. Moreover, resident Golgi enzymes gradually redistributed from
the juxtanuclear Golgi or Golgi ministacks to the ER in cells treated
with these PLA2 antagonists alone. Not only was ER-to-Golgi
transport of resident Golgi enzymes inhibited in cells treated with
these PLA2 antagonists, but transport of the vesicular
stomatitis virus G protein out of the ER was also prevented. These
results support a model of obligate retrograde recycling of Golgi
resident enzymes during nocodazole-induced ministack formation and
provide additional evidence that resident Golgi enzymes slowly and
constitutively cycle between the Golgi and ER.
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