Abstract
Taurine is known to function as a protectant against various stresses in animal cells. In order to utilize taurine as a compatible solute for stress tolerance of yeast, isolation of cDNA clones for genes encoding enzymes involved in biosynthesis of taurine was attempted. Two types of cDNA clones corresponding to genes encoding cysteine dioxygenase (CDO1 and CDO2) and a cDNA clone for cysteine sulfinate decarboxylase (CSD) were isolated from Cyprinus carpio. Deduced amino acid sequences of the two CDOs and that of CSD showed high similarity to those of CDOs and those of CSDs from other organisms, respectively. The coding regions of CDO1, CDO2, and CSD were subcloned into an expression vector, pESC-TRP, for Saccharomyces cerevisiae. Furthermore, to enhance the efficiency of synthesis of taurine in S. cerevisiae, a CDO–CSD fusion was designed and expressed. Expression of CDO and CSD proteins, or the CDO–CSD fusion protein was confirmed by Western blot analysis. HPLC analysis showed that the expression of the proteins led to enhancement of the accumulation level of hypotaurine, a precursor of taurine, rather than taurine. The yeast cells expressing corresponding genes showed tolerance to oxidative stress induced by menadione, but not to freezing–thawing stress.
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Abbreviations
- CDO:
-
Cysteine dioxygenase
- CSD:
-
Cysteine sulfinate decarboxylase
- DIG:
-
Digoxygenin
- NBT:
-
Nitro blue tetrazolium
References
Altschul SF, Gish W, Miller W, Myers EW, Lipman D (1990) Basic local alignment search tool. J Mol Biol 215:403–410
Bagley PJ, Hirschberger LL, Stipanuk MH (1995) Evaluation and modification of an assay procedure for cysteine dioxygenase activity: high-performance liquid chromatography method for measurement of cysteine sulfinate and demonstration of physiological relevance of cysteine dioxygenase activity in cysteine catabolism. Anal Biochem 227:40–48
Bidlingmeyer BA, Cohen SA, Tarvin TL (1984) Rapid analysis of amino acids using pre-column derivatization. J Chromatogr B 336:93–104
Bulow L, Mosbach K (1991) Multienzyme systems obtained by gene fusion. Trends Biotechnol 9:226–231
Chen Y, Foote RH, Brockett CC (1993) Effect of sucrose, trehalose, hypotaurine, taurine, and blood serum on survival of frozen bull sperm. Cryobiology 30:423–431
Daniels KM, Stipanuk MH (1982) The effect of dietary cysteine level on cysteine metabolism in rats. J Nutr 112:2130–2141
Dominy J, Eller S, Dawson R Jr (2004) Building biosynthetic schools: reviewing compartmentation of CNS taurine synthesis. Neurochem Res 29:97–103
Dominy JE, Hwang J, Guo S, Hirschberger LL, Zhang S, Stipanuk MH (2008) Synthesis of amino acid cofactor in cysteine dioxygenase is regulated by substrate and represents a novel post-translational regulation of activity. J Biol Chem 283:12188–12201
Hassan HM, Fridovich I (1979) Intracellular production of superoxide radical and of hydrogen peroxide by redox active compounds. Arch Biochem Biophys 196:385–395
Hayes KC (1985) Taurine requirement in primates. Nutr Rev 43:65–70
Heafield MT, Fearn S, Steventon GB, Waring RH, Williams AC, Sturman SG (1990) Plasma cysteine and sulphate levels in patients with motor neurone, Parkinson’s and Alzheimer’s disease. Neurosci Lett 110:216–220
Hoffman CS, Winston F (1987) A 10-minute DNA preparation from yeast efficiently releases autonomous plasmids for transformation of Escherichia coli. Gene 57:267–272
Honjoh K, Machida T, Nishi K, Matsuura K, Soli KW, Sakai T, Ishikawa H, Matsumoto K, Miyamoto T, Iio M (2007) Improvement of freezing and oxidative-stress tolerance in Saccharomyces cerevisiae by taurine. Food Sci Technol Res 13:145–154
Hosokawa Y, Matsumoto A, Oka J, Itakura H, Yamaguchi K (1990) Isolation and characterization of a cDNA for rat liver cysteine dioxygenase. Biochem Biophys Res Commun 168:473–478
Huxtable RJ (1992) Physiological actions of taurine. Physiol Rev 72:101–163
Ito H, Fukuda Y, Murata K, Kimura A (1983) Transformation of intact yeast cells treated with alkali cations. J Bacteriol 153:163–168
Kaisakia PJ, Jerkins AA, Goodspeed DC, Steele RD (1995) Cloning and characterization of rat cysteine sulfinic acid decarboxylase. Biochim Biophys Acta 1262:79–82
Kendall EJ, McKersie BD (1989) Free radical and freezing injury to cell membrane of winter wheat. Physiol Plant 76:86–94
Kyte J, Doolittle RF (1982) A simple method for displaying the hydropathic character of a protein. J Mol Biol 157:105–132
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T7. Nature 227:680–685
Mazur P (1970) Cryobiology: the freezing of biological systems. Science 168:353–367
McCann KP, Akbari MT, Williams AC, Ramsden DB (1994) Human cysteine dioxygenase type I: primary structure derived from base sequencing of cDNA. Biochim Biophys Acta 1209:107–110
Nakao M, Fushitani Y, Fujiki K, Nonaka M, Yano T (1998) Two diverged complement factor B/C2-like cDNA sequences from a teleost, the common carp (Cyprinus carpio). J Immunol 161:4811–4818
Park E, Park SY, Wang CH, Xu J, LaFauci G, Schuller-Levis G (2002) Cloning of murine cysteine sulfinic acid decarboxylase and its mRNA expression in murine tissues. Biochim Biophys Acta 1574:403–406
Perry TL, Kish SJ, Hansen S (1979) γ-Vinyl GABA: effects of chronic administration on the metabolism of GABA and other amino compounds in rat brain. J Neurochem 32:1641–1645
Reymond I, Sergeant A, Tappaz M (1996) Molecular cloning and sequence analysis of the cDNA encoding rat liver cysteine sulfinate decarboxylase (CSD). Biochim Biophys Acta 1307:152–156
Schaffer S, Azuma J, Takahashi K, Mozaffari M (2003) Why is taurine cytoprotective? Taurine 5. Adv Exp Med Biol 526:307–321
Seo HS, Koo YJ, Lim JY, Song JT, Kim CH, Kim JK, Lee JS, Choi YD (2000) Characterization of a bifunctional enzyme fusion of trehalose-6-phosphate synthetase and trehalose-6-phosphate phosphatase of Escherichia coli. Appl Environ Microbiol 66:2484–2490
Sherman F, Fink GR, Hickes JB (1986) Laboratory course manual for methods in yeast genetics. Cold Spring Harbor, NY
Shi XL, Flynn DC, Porter DW, Leonard SS, Valllyathan V, Castranova V (1997) Efficacy of taurine based compounds as hydroxyl radical scavengers in silica induced peroxidation. Ann Clin Lab Sci 27:365–374
Stintzing FC, Schieber A, Carle R (2001) Phytochemical and nutritional significance of cactus pear. Eur Food Res Technol 212:396–407
Stipanuk MH, Ueki I, Dominy JE Jr, Simmons CR, Hirschberger LL (2009) Cysteine dioxygenase: a robust system for regulation of cellular cysteine levels. Amino Acids 37:55–63
Takeuchi K, Toyohara H, Sakaguchi M (2000) A hyperosmotic stress-induced mRNA of carp cell encodes Na+- and Cl-dependent high affinity taurine transporter. Biochim Biophys Acta 1464:219–230
Towbin H, Stachelin T, Gorden J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some application. Proc Natl Acad Sci USA 76:4350–4354
Yanai H (2004) Statcel—the useful add-in software forms on Excel, 2nd edn. OMS, Tokyo
Yokoyama M, Takeuchi T, Park GS, Nakazoe J (2001) Hepatic cysteinesulfinate decarboxylase activity in fish. Aquac Res 32:220–261
Acknowledgments
This research was supported by a Grant-in-Aid for Scientific Research (C)(2) (no. 17580113) from the Japan Society for the Promotion of Science. The authors are grateful to Mr. Hsu-Ming Wen for the preparation of this manuscript.
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Honjoh, Ki., Matsuura, K., Machida, T. et al. Enhancement of menadione stress tolerance in yeast by accumulation of hypotaurine and taurine: co-expression of cDNA clones, from Cyprinus carpio, for cysteine dioxygenase and cysteine sulfinate decarboxylase in Saccharomyces cerevisiae . Amino Acids 38, 1173–1183 (2010). https://doi.org/10.1007/s00726-009-0328-6
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DOI: https://doi.org/10.1007/s00726-009-0328-6