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Mol Cell Biol, February 1998, p. 1125-1135, Vol. 18, No. 2
0270-7306/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Okadaic Acid Induces Selective Arrest of Protein Transport in the Rough Endoplasmic Reticulum and Prevents Export into COPII-Coated Structures

James G. Pryde,1 Theodora Farmaki,2 and John M. Lucocq2,*

Respiratory Medicine Unit, Department of Medicine (RIE), Rayne Laboratory, The University of Edinburgh Medical School, Edinburgh EH8 9AG,1 and Department of Anatomy and Physiology, The University of Dundee, Dundee DD1 4HN,2 Scotland, United Kingdom

Received 12 May 1997/Returned for modification 26 June 1997/Accepted 23 October 1997

Quantitative immunoelectron microscopy and subcellular fractionation established the site of endoplasmic reticulum (ER)-Golgi transport arrest induced by the phosphatase inhibitor okadaic acid (OA). OA induced the disappearance of transitional element tubules and accumulation of the anterograde-transported Chandipura (CHP) virus G protein only in the rough ER (RER) and not at more distal sites. The block was specific to the early part of the anterograde pathway, because CHP virus G protein that accumulated in the intermediate compartment (IC) at 15°C could gain access to Golgi stack enzymes. OA also induced RER accumulation of the IC protein p53/p58 via an IC-RER recycling pathway which was resistant to OA and inhibited by the G protein activator aluminium fluoride. The role of COPII coats in OA transport block was investigated by using immunofluorescence and cell fractionation. In untreated cells the COPII coat protein sec 13p colocalized with p53/p58 in Golgi-IC structures of the juxtanuclear region and peripheral cytoplasm. During OA treatment, p53/p58 accumulated in the RER but was excluded from sec 13p-containing membrane structures. Taken together our data indicate that OA induces an early defect in RER export which acts to prevent entry into COPII-coated structures of the IC region.

* Corresponding author. Mailing address: Medical Science Institute, University of Dundee, Dundee DD1 4HN, Scotland, United Kingdom. Phone: 01382 344973. Fax: 01382 345507. E-mail: jlucocq{at}bad.dundee.ac.uk.




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