Summary.
The cysteine biosynthesis pathway differs between plants and the yeast Saccharomyces cerevisiae. The yeast MET25 gene encoded to O-acetylhomoserine sulfhydrylase (AHS) catalyzed the reaction that form homocysteine, which later can be converted into cystiene. In vitro studies show that this enzyme possesses also the activity of O-acetyl(thiol)lyase (OASTL) that catalyzes synthesis of cysteine in plants. In this study, we generated transgenic tobacco plants expressing the yeast MET25 gene under the control of a constitutive promoter and targeted the yeast protein to the cytosol or to the chloroplasts. Both sets of transgenic plants were taller and greener than wild-type plants. Addition of SO2, the substrate of the yeast enzyme caused a significant elevation of the glutathione content in representative plants from each of the two sets of transgenic plants expressing the yeast gene. Determination of non-protein thiol content indicated up to four-folds higher cysteine and 2.5-fold glutathione levels in these plants. In addition, the leaf discs of the transgenic plants were more tolerant to toxic levels of sulphite, and to paraquat, an herbicide generating active oxygen species.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
References
J Antoniewski H Robichon-Szulmajster (1973) ArticleTitleBiosynthesis of methionine and its control in wild type and regulatory mutants of Saccharomyces cerevisiae. Biochimie 55 529–539 Occurrence Handle1:CAS:528:DyaE3sXlt1emsLw%3D Occurrence Handle4585174
D Becker E Kemper J Schell R Masterson (1992) ArticleTitleNew plant binary vectors with selectable markers located proximal to the left T-DNA border. Plant Mol Biol 20 1195–1197 Occurrence Handle1:CAS:528:DyaK3sXltlaqug%3D%3D Occurrence Handle1463855
A Blaszczyk R Brodzik A Sirko (1999) ArticleTitleIncreased resistance to oxidative stress in transgenic tobacco plants overexpressing bacterial serine acetyltransferase. Plant J 20 237–243 Occurrence Handle10.1046/j.1365-313x.1999.00596.x Occurrence Handle1:CAS:528:DyaK1MXotFait7c%3D Occurrence Handle10571883
P Buchner H Takahashi MJ Hawkesford (2004) ArticleTitlePlant sulphate transporters: co-ordination of uptake, intracellular and long-distance transport. J Exp Botany 55 1765–1773 Occurrence Handle10.1093/jxb/erh206 Occurrence Handle1:CAS:528:DC%2BD2cXntFaju7Y%3D
H Cherest D Thomas Y Surdin-Kerjan (1993) ArticleTitleCysteine biosynthesis in Saccharomyces cerevisiae occurs through the transsulphuration pathway which has been built up by enzyme recruitment. J Bacteriol 175 5366–5374 Occurrence Handle1:CAS:528:DyaK2cXhtVSgtb8%3D Occurrence Handle8366024
R D’Andrea Y Surdin-Kerjan G Pure H Cherest (1987) ArticleTitleMolecular genetics of met 17 and met 25 mutants of Saccharomyces cerevisiae: intragenic complementation between mutations of a single structural gene. Mol Gen Genet 207 165–170 Occurrence Handle1:CAS:528:DyaL2sXksFeiu78%3D Occurrence Handle3299001
JR Dominguez-Solis G Gutierrez-Alcala JM Vega LC Romero C Gotor (2001) ArticleTitleThe cytosolic O-acetylserine(thiol)lyase gene is regulated by heavy metals and can function in cadmium tolerance. J Biol Chem 276 31474 Occurrence Handle10.1074/jbc.M009574200 Occurrence Handle1:CAS:528:DC%2BD3MXmsVejtrk%3D
M Droux ML Ruffet R Douce D Job (1998) ArticleTitleInteractions between serine acetyltransferase and O-acetylserine(thiol)lyase in higher plants–structural and kinetic properties of the free and bound enzymes. Eur J Biochem 255 235–245 Occurrence Handle10.1046/j.1432-1327.1998.2550235.x Occurrence Handle1:CAS:528:DyaK1cXksFahsr4%3D Occurrence Handle9692924
D Gamrasni I Matityahu R Amir (2005) ArticleTitleAggregates formed as a result of the expression of yeast Met2 gene in transgenic tobacco plants, stimulate the production of stress-protective metabolites and increased the plants tolerance to heat stress. Mol Breeding 15 65–74 Occurrence Handle10.1007/s11032-004-2737-2 Occurrence Handle1:CAS:528:DC%2BD2MXhslyrsrk%3D
Y Hacham U Gofna R Amir (2003) ArticleTitleIn vivo analysis of various substrates utilized by cystathionine γ-synthase and O-acetylhomoserine sulfhydrylase in methionine biosynthesis. Mol Biol Evol 20 1513–1520 Occurrence Handle1:CAS:528:DC%2BD3sXntlantbw%3D Occurrence Handle12832650
K Harms P von Ballmoos C Brunold R Hofgen H Hesse (2000) ArticleTitleExpression of a bacterial serine acetyltransferase in transgenic potato plants leads to increased levels of cysteine and glutathione. Plant J 22 335–343 Occurrence Handle10.1046/j.1365-313x.2000.00743.x Occurrence Handle1:CAS:528:DC%2BD3cXkvFSksbc%3D Occurrence Handle10849350
R Hell R Jost O Berkowitz M Wirtz (2002) ArticleTitleMolecular and biochemical analysis of the enzymes of cysteine biosynthesis in the plant Arabidopsis thaliana. Amino Acids 22 245–257 Occurrence Handle10.1007/s007260200012 Occurrence Handle1:CAS:528:DC%2BD38XkvFOmu70%3D Occurrence Handle12083068
H Hesse V Nikiforova B Gakiere R Hoefgen (2004) ArticleTitleMolecular analysis and control of cysteine biosynthesis: integration of nitrogen and sulphur metabolism. J Exp Botany 55 1283–1292 Occurrence Handle1:CAS:528:DC%2BD2cXkvVSrsbo%3D
R Hofgen O Kreft L Willmitzer H Hesse (2001) ArticleTitleManipulation of thiol contents in plants. Amino Acids 20 291–299 Occurrence Handle1:CAS:528:DC%2BD3MXks1GjsrY%3D Occurrence Handle11354605
R Horsch B Fry NL Hoffmann D Eichholtz SG Rogers RT Fraley (1985) ArticleTitleA simple and general method for transferring genes into plants. Science 227 1229–1231 Occurrence Handle1:CAS:528:DyaL2MXhtFSjsLc%3D
CG Kawashima M Noji M Nakamura Y Ogra KT Suzuki K Saito (2004) ArticleTitleHeavy metal tolerance of transgenic tobacco plants over-expressing cysteine synthase. Biotechnol Lett 26 153–157 Occurrence Handle10.1023/B:BILE.0000012895.60773.ff Occurrence Handle1:CAS:528:DC%2BD2cXlt1ehsg%3D%3D Occurrence Handle15000484
UK Laemmli (1970) ArticleTitleCleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227 680–685 Occurrence Handle10.1038/227680a0 Occurrence Handle1:CAS:528:DC%2BD3MXlsFags7s%3D Occurrence Handle5432063
Liszewska F, Sirko A (2003) Analysis of transgenic tobacco lines expressing bacterial cysK gene encoding O-acetylserine(thiol)lyase A. In: Davidian J-C, Grill D, De Kok LJ, Stulen I, Hawkesford MJ, Schnung E, Rennerberg H (eds) Sulphur transport and assimilation in plants: regulation, interaction and signaling. Backhuys Publishers, Leiden, pp 269–271
F Liszewska A Blaszczyk A Sirko (2001) ArticleTitleModification of non-protein thiols contents in transgenic tobacco plants producing bacterial enzymes of cysteine biosynthesis pathway. Acta Biochimica Polonica 48 647–656 Occurrence Handle1:CAS:528:DC%2BD3MXovVGjtbc%3D Occurrence Handle11833773
T Leustek K Saito (1999) ArticleTitleSulfate transport and assimilation in plants. Plant Physiol 120 637–644 Occurrence Handle10.1104/pp.120.3.637 Occurrence Handle1:CAS:528:DyaK1MXks1amt7g%3D Occurrence Handle10398698
M Noji K Saito (2002) ArticleTitleMolecular and biochemical analysis of serine acetyltransferase and cysteine synthase towards sulfur metabolic engineering in plants. Amino Acids 22 231–243 Occurrence Handle10.1007/s007260200011 Occurrence Handle1:CAS:528:DC%2BD38XkvFOmu7w%3D Occurrence Handle12083067
M Noji M Saito M Nakamura M Aono H Saji K Saito (2001) ArticleTitleCysteine synthase overexpression in tobacco confers tolerance to sulfur-containing environmental pollutants. Plant Physiol 126 973–980 Occurrence Handle10.1104/pp.126.3.973 Occurrence Handle1:CAS:528:DC%2BD3MXlsVartb0%3D Occurrence Handle11457948
A Riemenschneider K Riedel R Hoefgen J Papenbrock H Hesse (2005) ArticleTitleImpact of reduced O-acetylserine(thiol)lyase isoform contents on potato plant metabolism. Plant Physiol 137 892–900 Occurrence Handle10.1104/pp.104.057125 Occurrence Handle1:CAS:528:DC%2BD2MXislOru7o%3D Occurrence Handle15728339
C Rochat JP Boutin (1991) ArticleTitleMetabolism of phloem-borne amino acids in maternal tissues of fruit of Nodulated or nitrate-fed pea plants (Pisum sativum L.). J Exp Bot 42 207–214 Occurrence Handle1:CAS:528:DyaK3MXhtl2qsrY%3D
K Saito (2000) ArticleTitleRegulation of sulfate transport and synthesis of sulfur-containing amino acids. Curr Opin Plant Biol 3 188–195 Occurrence Handle1:CAS:528:DC%2BD3cXktVOnsr0%3D Occurrence Handle10837270
K Saito M Kurosawa K Tatsuguchi Y Takagi I Murakoshi (1994) ArticleTitleModulation of cysteine biosynthesis in chloroplasts of transgenic tobacco overexpressing cysteine synthase [O-acetylserine(thiol)-lyase]. Plant Physiol 106 887–895 Occurrence Handle10.1104/pp.106.3.887 Occurrence Handle1:STN:280:ByqC38rht1w%3D Occurrence Handle7824657
O Shaul G Galili (1992) ArticleTitleIncreased lysine synthesis in tobacco plants that express high levels of bacterial dihydrodipicolinate synthase in their chloroplasts. Plant J 2 203–209 Occurrence Handle10.1111/j.1365-313X.1992.00203.x Occurrence Handle1:CAS:528:DyaK38XkvVKqtLk%3D
A Sirko A Blaszczyk F Liszewska (2004) ArticleTitleOverproduction of SAT and/or OASTL in transgenic plants: a survey of effects. J Exp Botany 55 1881–1888 Occurrence Handle10.1093/jxb/erh151 Occurrence Handle1:CAS:528:DC%2BD2cXntFaisbs%3D
D Thomas Y Surdin-Kerjan (1997) ArticleTitleMetabolism of sulfur amino acids in Saccharomyces cerevisiae. Microbiol Mol Biol Rev 61 503–532 Occurrence Handle1:CAS:528:DyaK2sXotVymsbc%3D Occurrence Handle9409150
M Vanoni M Vai L Popolo L Alberghina (1983) ArticleTitleStructural heterogeneity in populations of the budding yeast Saccharomyces cerevisiae. J Bacteriol 156 1282–1291 Occurrence Handle1:CAS:528:DyaL2cXitVarsA%3D%3D Occurrence Handle6358196
M Wirtz R Hell (2003) ArticleTitleProduction of cysteine for bacterial and plant biotechnology: application of cysteine feedback-insensitive isoforms of serine acetyltransferase. Amino Acids 24 195–203 Occurrence Handle1:CAS:528:DC%2BD3sXhslGjs7Y%3D Occurrence Handle12624753
S Yamagata K Takeshima N Naiki (1975) ArticleTitleO-acetylserine and O-acetylhomoserine sulfhydrylase of yeast; studies with methionine auxotrophs. J Biochem (Tokyo) 77 1029–1036 Occurrence Handle1:CAS:528:DyaE2MXksFGlsr8%3D
S Yamagata M Isaji K Nakamura S Fujisaki K Doi S Bawden R D’Andrea (1994) ArticleTitleOverexpression of the Saccharomyces cerevisiae MET17/MET25 gene in Escherichia coli and comparative characterization of the product with O-acetylserine. O-acetylhomoserine sulfhydrylase of the yeast. Appl Microbiol Biotechnol 42 92–99 Occurrence Handle1:CAS:528:DyaK2MXktVSltb0%3D Occurrence Handle7765825
S Youssefian M Nakamura H Sano (1993) ArticleTitleTobacco plants transformed with the O-acetylserine(thiol)lyase gene of wheat are resistant to toxic levels of hydrogen sulphide gas. Plant J 4 759–769 Occurrence Handle10.1046/j.1365-313X.1993.04050759.x Occurrence Handle1:CAS:528:DyaK2cXitFSht7o%3D Occurrence Handle8275097
S Youssefian M Nakamura E Orudgev N Kondo (2001) ArticleTitleIncreased cysteine biosynthesis capacity of transgenic tobacco overexpressing an O-acetylserine(thiol)lyase modifies plant responses to oxidative stress. Plant Physiol 126 1001–1111 Occurrence Handle10.1104/pp.126.3.1001 Occurrence Handle1:CAS:528:DC%2BD3MXlsVartbg%3D Occurrence Handle11457951
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Matityahu, I., Kachan, L., Bar Ilan, I. et al. Transgenic tobacco plants overexpressing the Met25 gene of Saccharomyces cerevisiae exhibit enhanced levels of cysteine and glutathione and increased tolerance to oxidative stress. Amino Acids 30, 185–194 (2006). https://doi.org/10.1007/s00726-005-0250-5
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00726-005-0250-5