Mannose-dependent Endoplasmic Reticulum (ER)-Golgi Intermediate Compartment-53-mediated ER to Golgi Trafficking of Coagulation Factors V and VIII*
- Micheline Moussalli‡§,
- Steven W. Pipe¶§,
- Hans-Peter Hauri‖,
- William C. Nichols**,
- David Ginsburg‡‡§§ and
- Randal J. Kaufmanদ,166
- From the Departments of ¶Pediatrics,‡Medicine, §§Human Genetics, and ¶¶Biological Chemistry, the‡Howard Hughes Medical Institute, University of Michigan School of Medicine, Ann Arbor, Michigan 48109, the **Division of Human Genetics, Children's Hospital Medical Center, Cincinnatti, Ohio, and the ‖Department of Pharmacology/Neurobiology, Biozentrum, University of Basel, Basel CH-4056, Switzerland
Abstract
The endoplasmic reticulum-Golgi intermediate compartment (ERGIC) is the site of segregation of secretory proteins for anterograde transport, via packaging into COPII-coated transport vesicles. ERGIC-53 is a homo-hexameric transmembrane lectin localized to the ERGIC that exhibits mannose-selective properties in vitro. Null mutations in ERGIC-53 were recently shown to be responsible for the autosomal recessive bleeding disorder, combined deficiency of coagulation factors V and VIII. We have studied the effect of defective ER to Golgi cycling by ERGIC-53 on the secretion of factors V and VIII. The secretion efficiency of factor V and factor VIII was studied in a tetracycline-inducible HeLa cell line overexpressing a wild-type ERGIC-53 or a cytosolic tail mutant of ERGIC-53 (KKAA) that is unable to exit the ER due to mutation of two COOH-terminal phenylalanine residues to alanines. The results show that efficient trafficking of factors V and VIII requires a functional ERGIC-53 cycling pathway and that this trafficking is dependent on post-translational modification of a specific cluster of asparagine (N)-linked oligosaccharides to a fully glucose-trimmed, mannose9 structure.
Footnotes
-
↵* Portions of this work were supported by National Institutes of Health Grant HL5734601A1 (to D. G.) and Grant HL5217302 (to R. J. K.).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.
-
↵§ These authors contributed equally to the work.
-
↵166 To whom correspondence should be addressed: 1150 W. Medical Ctr. Dr., MSRB II, Rm. 4570, Ann Arbor, MI 48109. Tel.: 313-763-9037; Fax: 313-763-9323; E-mail: kaufmanr@umich.edu.
- Abbreviations:
- ERGIC
-
endoplasmic reticulum-Golgi intermediate compartment
- BDD
-
B domain deleted
- WT
-
wild-type
- CST
-
castanospermine
- DMJ
-
deoxymannojirimycin
-
- Received August 16, 1999.
- The American Society for Biochemistry and Molecular Biology, Inc.
