Nordihydroguaiaretic Acid Blocks Protein Transport in the Secretory Pathway Causing Redistribution of Golgi Proteins into the Endoplasmic Reticulum*
- From the Department of Biochemistry and ‡Radioisotope Laboratory, Fukuoka University School of Medicine, Jonan-ku, Fukuoka 814-80, Japan
Abstract
We have investigated the effect of nordihydroguaiaretic acid (NDGA), an inhibitor of lipoxygenase, on the intracellular protein transport and the structure of the Golgi complex. Pulse-chase experiments and immunoelectron microscopy showed that NDGA strongly inhibits the transport of newly synthesized secretory proteins to the Golgi complex resulting in their accumulation in the endoplasmic reticulum (ER). Despite their retention in the ER, oligosaccharides of secretory and ER-resident proteins were processed to endoglycosidase H-resistant forms, raising the possibility that oligosaccharide-processing enzymes are redistributed from the Golgi to the ER. Morphological observations further revealed that α-mannosidase II (a cis/medial-Golgi marker), but not TGN38 (a trans-Golgi network marker), rapidly redistributes to the ER in the presence of NDGA, resulting in the disappearance of the characteristic Golgi structure. Upon removal of the drug, the Golgi complex was reassembled into the normal structure as judged by perinuclear staining of α-mannosidase II and by restoration of the secretion. These effects of NDGA are quite similar to those of brefeldin A. However, unlike brefeldin A, NDGA did not cause a dissociation of β-coatomer protein, a subunit of coatomer, from the Golgi membrane. On the contrary, NDGA exerted the stabilizing effect on β-coatomer protein/membrane interaction against the dissociation caused by brefeldin A and ATP depletion. Taken together, these results indicate that NDGA is a potent agent disrupting the structure and function of the Golgi complex with a mechanism different from those known for other drugs reported so far.
Footnotes
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↵* This work was supported by grants from the Ministry of Education, Science, Sports, and Culture of Japan, the Japan Private School of Promotion Foundation, the Japan Science and Technology Corporation, the Naito Foundation, and The Central Research Institute of Fukuoka University.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked “advertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
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↵§ To whom correspondence should be addressed: Tel.: 81-92-801-1011 (ext. 3250); Fax: 81-92-864-3865; E-mail:mm031321{at}cc.fukuoka-u.ac.jp.
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↵1 The abbreviations used are: ER, endoplasmic reticulum; ARF, ADP-ribosylation factor; BFA, brefeldin A; C3, third component of complement; CGN, cis-Golgi network; COP, coatomer protein complex; DPP IV, dipeptidyl-peptidase IV; endo H, endo-β-hexosaminidase H; Man II, α-mannosidase II; NDGA, nordihydroguaiaretic acid; α1-PI, α1-protease inhibitor; TGN, trans-Golgi network; PAGE, polyacrylamide gel electrophoresis; PBS, phosphate-buffered saline; GTPγS, guanosine 5′-3-O-(thio)triphosphate.
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- Received June 19, 1997.
- Revision received October 17, 1997.
- The American Society for Biochemistry and Molecular Biology, Inc.
