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J Biol Chem, Vol. 275, Issue 19, 14494-14500, May 12, 2000

The Phosphorylation State and Expression of Soybean BiP Isoforms Are Differentially Regulated following Abiotic Stresses*

Júlio Cézar M. CascardoDagger §, Raul S. Almeida||, Reginaldo A. A. Buzeli||, Sônia M. B. CarolinoDagger §, Wagner C. OtoniDagger , and Elizabeth P. B. Fontes**

From the Dagger  Departamento de Biologia Vegetal and  Departamento de Bioquímica e Biologia Molecular/Instituto de Biotecnologia Aplicado à Agropecuária, Universidade Federal de Viçosa, 36571.000 Viçosa Minas Gerais, Brazil

The mammalian BiP is regulated by phosphorylation, and it is generally accepted that its unmodified form constitutes the biologically active species. In fact, the glycosylation inhibitor tunicamycin induces dephosphorylation of mammalian BiP. The stress-induced phosphorylation state of plant BiP has not been examined. Here, we demonstrated that soybean BiP exists in interconvertible phosphorylated and nonphosphorylated forms, and the equilibrium can be shift to either direction in response to different stimuli. In contrast to tunicamycin treatment, water stress condition stimulated phosphorylation of BiP species in soybean cultured cells and stressed leaves. Despite their phosphorylation state, we demonstrated that BiP isoforms from water-stressed leaves exhibit protein binding activity, suggesting that plant BiP functional regulation may differ from other eukaryotic BiPs. We also compared the induction of the soybean BiP gene family, which consists of at least four members designated soyBiPA, soyBiPB, soyBiPC, and soyBiPD, by tunicamycin and osmotic stress. Although all soybean BiP genes were induced by tunicamycin, just the soyBiPA RNA was up-regulated by osmotic stress. In addition, these stresses promoted BiP induction with different kinetics and acted synergistically to increase BiP accumulation. These results suggest that the soybean BiP gene family is differentially regulated by abiotic stresses through distinct signaling pathways.

* This research was supported by the Brazilian Government Agency, Financiadora de Estudos e Projetos/Fundo Nacional para Desenvolvimento Científico e Tecnológico, Grant 64.94.0113.00 (to E. P. B. F.).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.

§ Recipients of graduate Conselho Nacional de Desenvolvimento Científico e Tecnológico fellowships from the Brazilian Government.

|| Recipients of Fundação de Amparo à Pesquisa do Estado de Minas Gerais scholarships from the Minas Gerais State, Brazil.

** To whom correspondence should be addressed: DBB/BIOAGRO, Universidade Federal de Viçosa, Av PH Rolfs s/n, 36571.000 Viçosa MG, Brazil. Tel.: 01-15-53-1899-2949; Fax: 01-15-53-1899-2864; E-mail: bbfontes@mail.ufv.br.

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