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Molecular Cloning and Expression of Sequence Variants of Manganese Superoxide Dismutase Genes from Wheat
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 Title & Authors
Molecular Cloning and Expression of Sequence Variants of Manganese Superoxide Dismutase Genes from Wheat
Baek, Kwang-Hyun; Skinner, Daniel Z.;
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Reactive oxygen species (ROS) are very harmful to living organisms due to the potential oxidation of membrane lipids, DNA, proteins, and carbohydrates. transformed E.coli strain QC 871, superoxide dismutase (SOD) double-mutant, with three sequence variant MnSOD1, MnSOD2, and MnSOD3 manganese superoxide dismutase (MnSOD) gene isolated from wheat. Although all QC 871 transformants grown at expressed mRNA of MnSOD variants, only MnSOD2 transformant had functional SOD activity. MnSOD3 expressed active protein when grown at , however, MnSOD1 did not express functional protein at any growing and induction conditions. The sequence comparison of the wheat MnSOD variants revealed that the only amino acid difference between the sequence MnSOD2 and sequences MnSOD1 and 3 is phenylalanine/serine at position 58 amino acid. We made MnSOD2S58F gene, which was made by altering the phenylalaine to serine at position 58 in MnSOD2. The expressed MnSOD2S58F protein had functional SOD activity, even at higher levels than the original MnSOD2 at all observed temperatures. These data suggest that amino acid variation can result in highly active forms of MnSOD and the MnSOD2S58F gene can be an ideal target used for transforming crops to increase tolerance to environmental stresses.
Environmental stress;Escherichia coli;Expression;Manganese superoxide dismutase;Multigene family;
 Cited by
Production of reactive oxygen species by freezing stress and the protective roles of antioxidant enzymes in plants, Journal of Agricultural Chemistry and Environment, 2012, 01, 01, 34  crossref(new windwow)
Allen, S.P., Polazzi, J.O., Gierse, J.K., Easton, A.M., 1992.Two novel heat shock genes encoding proteins produced in response to heterologous protein expression in E. coli, J. Bacteriol. 174, 6938-6947.

Allen, R.D., 1995. Dissection of oxidative stress tolerance using transgenic plants, Plant Physiol. 107, 1049-1054.

Baek, K.-H., Skinner, D.Z., 2003. Alteration of antioxi-dant enzyme gene expression during cold acclimation of near-isogenic wheat lines, Plant Sci. 165, 1221-1227. crossref(new window)

Baek, K.-H., Skinner, D.Z., 2006. Differential expression of manganese superoxide dismutase sequence variants in near isogenic lines of wheat during cold accli-mation, Plant Cell Rep. 25: 223-230. crossref(new window)

Basu, U., Good, A.G., Taylor, G.J., 2001. Transgenic Brassica napus plants overexpressing aluminum-induced mitochondrial manganese superoxide dismutase cDNA are resistant to aluminum, Plant, Cell, and Environ. 24, 1269-1278.

Beauchamp, C.O., Fridovich, I., 1971. Superoxide dismutase: improved assays and an assay applicable to acrylamide gels, Anal. Biochem. 44, 276-287. crossref(new window)

Borgstahl G.E.O., Parge H.E., Hickey M.J., Johnson M.J., Boissinot M., Hallewell R.A., Lepock J.R., Cabelli D.E., Tainer J.A, 1996. Human mitochon-drial manganese superoxide dismutase polymorphic variant Ile58Thr reduces activity by destabilizing the tetrameric interface, Biochemistry 35: 4287-4297. crossref(new window)

Bowler, C., Slooten, L., Vandenbranden, S., De Rycke, R., Botterman, J., Sybesma, C., Van Montague, M., Inze, D., 1991. Manganese superoxide dismutase can reduce cellular damage mediated by oxygen radicals in transgenic plants, EMBO J. 10, 1723-1732.

Bowler, C., Montagu, M.V., Inze, D., 1992. Superoxide dismutase and stress tolerance, Annu. Rev. Plant Physiol. Plant Mol. Biol. 43, 83-116. crossref(new window)

Breusegem, F. , Slooten, L., Stassart, J.M., Botterman, J., Moens, T., Montagu, M., Inze, D., 1999. Effects of overproduction of tobacco MnSOD in maize chloroplasts on foliar tolerance to cold and oxidative stress, J. Exp. Bot. 50, 71-78. crossref(new window)

Carlioz, A., Touati, D., 1986. Isolation of superoxide dismutase mutants in Escherichia coli: Is superoxide dismutase necessary for aerobic life?, EMBO J. 5, 623-630.

Chen, C.-N., Pan, S.-M., 1996. Assay of superoxide dismutase activity by combining electrophoresis and densitometry, Bot. Bull.Acad. Sin. 37, 107-111.

Fischer, B., Sumner, L., Goodenough, P., 1993. Isolation, renaturation, and formation of disulfide bonds of eukaryotic proteins expressed in Escherichia coli as inclusion bodies. Biotechnol. Bioeng. 41, 3-13. crossref(new window)

Fucci, L., Oliver, C.N., Coon, M.J., Stadtman, E.R.,1983. Inactivation of metabolic enzymes by mixed-function oxidation reaction: possible implication in protein turnover and ageing, Proc. Natl. Acad. Sci. U.S.A. 80, 1521-1525. crossref(new window)

Giannopolitis, C.N., Ries, K., 1977. Superoxide dis-mutase I. Occurrence in higher plants, Plant Physiol. 59, 309-314. crossref(new window)

Greenleaf, W.B., Perry, J.J., Hearn, A.S., Cabelli, D.E., Lepock, J.R., Stroupe, M.E., Tainer, J.A., Nick, H.S., Silverman, D.N. 2004. Role of hydrogen bonding in the active site of human manganese superoxide dismutase, Biochemistry 43, 7038-7045. crossref(new window)

Halliwell, B., Gutteridge, J.M.C., 2007. Free radicals in biology and medicine, Fourth ed. Oxford Unizver-sity Press, New York, pp 262-282.

Houot, V., Etienne, P., Petitot, A.S., Barbier, S., Blein, J.P., Suty, L., 2001. Hydrogen peroxide induces programmed cell death features in cultured tobacco BY-2 cells, in a dose-dependent manner, J. Exp. Bot. 52, 1721-1730. crossref(new window)

Kendall, E.J., McKersie, B.D., 1989. Free radical and freezing injury to cell membranes of winter wheat, Physiol. Plant. 76, 86-94. crossref(new window)

Kim, A., Joseph, S., Khan, A., Epstein, C.J., Sobel, R., Huang, T.T., 2010. Enhanced expression of mito-chondrial superoxide dismutase leads to prolonged in vivo cell cycle progression and up-regulation of mitochondrial thioredoxin, Free. Radic. Biol. Med. doi:10.1016/j.freeradbiomed.2010.02.028. crossref(new window)

Laemmli, U.K.,1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4, Nature 227, 680-685. crossref(new window)

Larkin, M.A., Blackshields, G., Brown, N.P., Chenna, R., McGettigan, P.A., McWilliam, H., Valentin,F., Wallace, I.M., Wilm, A., Lopez, R., Thompson, J.D., Gibson, T.J., Higgins, D.G., 2007. ClustalW and ClustalX version 2, Bioinformatics 23, 2947-2948. crossref(new window)

Lee, N., 1980. Molecular aspects of ara regulation, in: Miller, J.H., Reznikoff, W.S. (Eds) The Operon, Cold Spring Harbor, N.Y.: Cold Spring Harbor Laboratory, pp. 389-410.

Lee, N., Francklyn, C., Hamilton, E.P., 1987. Arabinose-Induced Binding of AraC Protein to aria Activates the araBAD Operon Promoter, Proc. Natl. Acad. Sci. U.S.A. 84, 8814-8818. crossref(new window)

McCord, J.M., Fridovich, I., 1969. Superoxide dismutase, an enzymic function for Erythrocuprein (Hemo-cuprein), J. Biol. Chem. 244, 6049-6055.

McKersie, B.D., Chen, Y.R., De Beus, M., Bowler, S.R., Inze, D., D’Halluin, K., Botterman, J., 1993. Superoxide dismutase enhances tolerance of freezing stress in transgenic alfalfa (Medicago sativa L.), Plant Physiol. 103, 1155-1163. crossref(new window)

Miao, Z., Gaynor, J.J., 1993. Molecular cloning, characteri-zation and expression of Mn-superoxide dismutase from the rubber tree (Hevea brasiliensis), Plant Mol. Biol. 23, 267-277. crossref(new window)

Rubio, M.C., Ramos, J., Webb, K.J., Minchin, F.R., Gonzalez, E., Arrese-Igor, C., Becana, M., 2001. Expression studies of superoxide dismutases in nodules and leaves of transgenic alfalfa reveal abundance of iron-containing isozymes, posttrans-lational regulation, and compensation of isozyme activities, Mol. Plant Microbe In. 14, 1178-1188. crossref(new window)

Rudolph, R., Lilie, H., 1996. In vitro folding of inclusion body proteins, FASEB J. 10, 49-56.

Sambrook, J., Russell, D.W., 2001. Molecular cloning: A laboratory manual, third ed. Cold Spring Laboratory, New York

Sato, I., Zu, J., Nishikawa, S., Kashimura, N., 1993. Depolymerization of hyaluronic acid by D-fructose 6-phosphate, Biosci. Biotechnol. Biochem. 57, 2005-2009. crossref(new window)

Schrank, I.S., Sims, P.F.G., Oliver, S.G., 1988. Functional expression of the yeast Mn-superoxide dismutase gene in Escherichia coli requires deletion of the signal peptide sequence, Gene 73, 121-130. crossref(new window)

Storlie, E.W., Allan, R.E., Walker-Simmons, M.K., 1998. Effect of the Vrn1-Fr1 interval on cold hardiness levels in near isogenic wheat lines, Crop Sci. 38, 483-488. crossref(new window)

Streller, S., Kromer, S., Wingsle, G., 1994. Isolation and purification of mitochondrial Mn-superoxide dismutase from the gymnosperm Pinus sylvestris L, Plant Cell Physiol. 35, 859-867.

Wang, Y.C., Qu, G, Z., Li, H.Y., Wu, Y.J., Wang, C., Liu, G.F.,Yang, C.P., 2010. Enhanced salt tolerance of transgenic poplar plants expressing a manganese superoxide dismutase from Tamarix androssowii, Mol. Biol. Rep. 37, 1119-1124. crossref(new window)

Wilder, M.S., Mass, A., 1990. Composition for pre-venting or alleviating skin irritation by formulations containing superoxide dismutase, U.S. Patent 4,957, 740, 1-12.

Wu, G., Shortt, B.J., Lawrence, E.B., Fitzsimmons, J.L.K.C., Levine, E.B., Raskin, I., Shah, D.M., 1997. Activation of host defense mechanisms by elevated production of $H_2O_2$ in transgenic plants, Plant Physiol. 115, 427-435.

Wu, G., Wilen, R.W., Robertson, A.J., Gusta, L.V., 1999. Isolation, chromosome localization, and differential expression of mitochondrial manganese superoxide dismutase and chloroplastic copper/Zinc superoxide dismutase genes in wheat, Plant Physiol. 120, 513-520. crossref(new window)

Xie, Y., Liu, Y., Meng, M., Chen, L., Zhu, Z., 2003. Isolation and identification of a super strong plant promoter from cotton leaf curl Multan virus, Plant Mol. Biol. 53, 1-14. crossref(new window)

Zelko, I.N., Mariani, T.J., Folz, R.J., 2002. Superoxide dismutase multigene family: A comparison of CuZn-SOD (SOD1), Mn-SOD (SOD2), and EC-SOD (SOD3) gene structures, evolution, and expression. Free Radic. Biol. Med. 33, 337-349. crossref(new window)

Zhu, D., Scandalios, J.G., 1993. Maize mitochondrial manganese superoxide dismutases are encoded by a differentially expressed multigene family, Proc. Natl. Acad. Sci. U.S.A. 90, 9310-9314. crossref(new window)