Abstract
The gene HvNHX3 encoding a new isoform of vacuolar Na+/H+-antiporter was identified in barley. This gene is expressed in roots and leaves of barley seedlings, and it encodes a protein consisting of 541 amino acid residues with pre-dicted molecular weight 59.7 kDa. It was found that by its amino acid sequence HvNHX3 is closest to the Na+/H+-antiporter HbNHX1 of wild type from Hordeum brevisibulatum that grows on salt-marsh (solonchak) soils (95% homology). The expression of HvNHX3 during salt stress is increased several-fold in roots and leaves of barley seedlings. At the same time, the amount of HvNHX3 protein in roots does not change, but in leaves it increases significantly. It was shown using HvNHX3 immunolocalization in roots that this protein is present in all tissues, but in control plants it was clustered and in experimental plants after salt stress it was visualized as small granules. It has been proposed that HvNHX3 is converted into active form during declusterization. The conversion of HvNHX3 into its active form along with its quantitative increase in leaves during salt stress activates Na+/H+-exchange across the vacuolar membrane and Na+ release from cytoplasm, and, as a consequence, an increase of salt stress tolerance.
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Abbreviations
- a.a.:
-
amino acid residue
- CBD:
-
cellulose-binding domain
- DTT:
-
dithiothreitol
- PMSF:
-
phenylmethylsulfonyl fluoride
- RT-PCR:
-
reverse transcription polymerase chain reaction
References
Niu, X., Bressan, R. A., Hasegawa, P. M., and Pardo, J. M. (1995) Plant Physiol., 109, 735–742.
Qiu, Q. S., Barkla, B. J., Vera-Estrella, R., Zhu, J.-K., and Schumaker, K. (2003) Plant Physiol., 132, 1041–1052.
Apse, M. P., Aharon, G. S., Snedden, W. A., and Blumwald, E. (1999) Science, 285, 1256–1258.
Yamaguchi, T., Fukuda-Tanaka, S., Inagaki, Y., Saito, N., Yonekura-Sakakibara, K., Tanaka, Y., Kusumi, T., and Iida, S. (2001) Plant Cell Physiol., 142, 451–461.
Apse, M. P., Sottosanto, J., and Blumwald, E. (2003) Plant J., 36, 229–239.
Brett, C. L., Tukaye, D. N., Mukherjee, S., and Rao, R. (2005) Mol. Biol. Cell, 16, 1396–1405.
Wu, Y. Y., Chen, G. D., Meng, Q. W., and Zheng, C. C. (2004) Plant Cell Physiol., 45, 600–607.
Saqib, M., Zorb, C., Rengel, Z., and Schubert, S. (2005) Plant Sci., 169, 959–965.
Yamaguchi, T., and Blumwald, E. (2005) Trends Plant Sci., 12, 615–620.
Padan, E., Venturi, M., Gerchman, Y., and Dover, N. (2001) Biochim. Biophys. Acta, 1505, 144–157.
Nass, R., and Rao, R. (1998) J. Biol. Chem., 273, 21054–21060.
Orlowski, J., and Grinstein, S. (1997) J. Biol. Chem., 272, 22373–22376.
Nakamura, N., Tanaka, S., Teko, Y., Mitsui, K., and Kanazawa, H. (2005) J. Biol. Chem., 280, 1561–1572.
Yokoi, S., Quintero, F. J., Cubero, B., Ruiz, M. T., Bressan, R. A., Hasegawa, P. M., and Pardo, J. M. (2002) Plant J., 30, 529–539.
Zorb, C., Noll, A., Karl, S., Leib, K., Yan, F., and Schubert, S. (2005) J. Plant Physiol., 162, 55–66.
Venema, K., Belver, A., Marin-Manzano, M. C., Rodriguez-Rosales, M. P., and Donairo, J. P. (2003) J. Biol. Chem., 278, 22453–22459.
Ohnishi, M., Fukada-Tanaka, S., Hoshino, A., Takada, J., Inagaki, Y., and Iida, S. (2005) Plant Cell Physiol., 46, 259–267.
Fukuda, A., Chiba, K., Maeda, M., Nakamura, A., Maeshima, M., and Tanaka, Y. (2004) J. Exp. Bot., 397, 585–594.
Vasekina, A. V., Ershov, P. V., Reshetova, O. S., Tikhonova, T. V., Lunin, V. G., Trofimova, M. S., and Babakov, A. V. (2005) Biochemistry, 70, 100–107.
Ershov, P. V., Vasekina, A. V., Voblikova, V. D., Taranov, V. V., Roslyakova, T. V., and Babakov, A. V. (2007) Fiziol. Rast., 54, 22–30.
Maeshima, M., and Yoshida, S. (1989) J. Biol. Chem., 264, 20068–20073.
Carlemaln, E., Garavito, R. M., and Villiger, W. (1982) J. Microsc., 126, 123–124.
Putney, L. K., Denker, S. P., and Barber, D. L. (2002) Annu. Rev. Pharmacol. Toxicol., 42, 527–552.
Danilova, M. F. (1974) Structural Basis of the Root Absorption [in Russian], Nauka, Moscow.
Brett, C. L., Donowitz, M., and Rao, R. (2005) Am. J. Physiol. Cell Physiol., 288, 223–239.
Venema, K., Quintero, F. J., Pardo, J. M., and Donaire, J. P. (2002) J. Biol. Chem., 277, 2413–2418.
Yamaguchi, T., Apse, M. P., Shi, H., and Blumwald, E. (2003) Proc. Natl. Acad. Sci. USA, 100, 12510–12515.
Lu, S. Y., Jing, Y. X., Shen, S. H., Zhao, H. Y., Ma, L. Q., Zhou, X. J., Ren, Q., and Li, Y. F. (2007) J. Integr. Plant Biol., 47, 343–349.
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Original Russian Text © T. V. Roslyakova, E. M. Lazareva, N. V. Kononenko, A. V. Babakov, 2009, published in Biokhimiya, 2009, Vol. 74, No. 5, pp. 675–684.
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Roslyakova, T.V., Lazareva, E.M., Kononenko, N.V. et al. New isoform HvNHX3 of vacuolar Na+/H+-antiporter in barley: Expression and immunolocalization. Biochemistry Moscow 74, 549–556 (2009). https://doi.org/10.1134/S0006297909050101
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DOI: https://doi.org/10.1134/S0006297909050101