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Originally published In Press as doi:10.1074/jbc.M003061200 on May 18, 2000

J. Biol. Chem., Vol. 275, Issue 31, 23685-23692, August 4, 2000
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Endoplasmic Reticulum Oxidoreductin 1-Lbeta (ERO1-Lbeta ), a Human Gene Induced in the Course of the Unfolded Protein Response*,

Massimiliano PaganiDagger §, Marco FabbriDagger §, Cristina BenedettiDagger , Anna FassioDagger , Stefania PilatiDagger , Neil J. Bulleid||, Andrea CabibboDagger , and Roberto SitiaDagger **

From the Dagger  Department of Molecular Pathology and Medicine, DIBIT-San Raffaele Scientific Institute, Via Olgettina 58, 20132 Milano, Italy and the || School of Biological Sciences, University of Manchester, 2.205 Stopford Building, Oxford Road, Manchester M13 9PT, United Kingdom

Oxidative conditions must be generated in the endoplasmic reticulum (ER) to allow disulfide bond formation in secretory proteins. A family of conserved genes, termed ERO for ER oxidoreductins, plays a key role in this process. We have previously described the human gene ERO1-L, which complements several phenotypic traits of the yeast thermo-sensitive mutant ero1-1 (Cabibbo, A., Pagani, M., Fabbri, M., Rocchi, M., Farmery, M. R., Bulleid, N. J., and Sitia, R. (2000) J. Biol. Chem. 275, 4827-4833). Here, we report the cloning and characterization of a novel human member of this family, ERO1-Lbeta . Immunofluorescence, endoglycosidase sensitivity, and in vitro translation/translocation assays reveal that the products of the ERO1-Lbeta gene are primarily localized in the ER of mammalian cells. The ability to allow growth at 37 °C and to alleviate the "unfolded protein response" when expressed in ero1-1 cells indicates that ERO1-Lbeta is involved also in generating oxidative conditions in the ER. ERO1-L and ERO1-Lbeta display different tissue distributions. Furthermore, only ERO1-Lbeta transcripts are induced in the course of the unfolded protein response. Our results suggest a complex regulation of ER redox homeostasis in mammalian cells.

* This work was supported in part through grants from Associazione per la Ricerca sul Cancro, Consiglio Nazionale delle Ricerche (Target Project on Biotechnology, Grant CNR 97.01296.PF49; 5% Biotechnology, Grant CNR 98.00393.PF31), Ministero della Sanità (RF 9853), and Biotechnology and Biological Sciences Research Council (Grant 34/C09198).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.

The nucleotide sequence(s) reported in this paper has been submitted to the GenBankTM/EMBL Data Bank with accession number(s) AF252538.

The on-line version of this article (available at http://www.jbc.org) contains supplementary quantitative data on the tissue distribution of ER01-Lalpha , ER01-Lbeta , and ubiquitin obtained by PhosphorImager (Molecular Probes). The ratios between the different transcripts in the various tissues are also reported.

§ These authors contributed equally to this work.

Present address: Dept. of Applied and Molecular Ecology, Waite Campus, Glen Osmond, South Australia 5064, Australia.

** To whom correspondence should be addressed: DIBIT-HSR Via Olgettina 58, 20132 Milano, Italy. Tel.: +39-02-2643-4763; Fax: +39-02-2643-4723; E-mail: r.sitia@hsr.it.

Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
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