Expression analysis of a novel pyridoxal kinase messenger RNA splice variant, PKL, in oil rape suffering abiotic stress and phytohormones
Shunwu Yu
Shanghai Agrobiological Gene Center, Shanghai Academy of Agricultural Sciences, Germsperm Resources Division (Shanghai), National Key Laboratory of Crop Genetic Improvement, Shanghai 201106, China
Search for more papers by this authorCorresponding Author
Lijun Luo
Shanghai Agrobiological Gene Center, Shanghai Academy of Agricultural Sciences, Germsperm Resources Division (Shanghai), National Key Laboratory of Crop Genetic Improvement, Shanghai 201106, China
*Corresponding author: Tel, 86-21-52230526; Fax, 86-21-62204010; E-mail, [email protected]Search for more papers by this authorShunwu Yu
Shanghai Agrobiological Gene Center, Shanghai Academy of Agricultural Sciences, Germsperm Resources Division (Shanghai), National Key Laboratory of Crop Genetic Improvement, Shanghai 201106, China
Search for more papers by this authorCorresponding Author
Lijun Luo
Shanghai Agrobiological Gene Center, Shanghai Academy of Agricultural Sciences, Germsperm Resources Division (Shanghai), National Key Laboratory of Crop Genetic Improvement, Shanghai 201106, China
*Corresponding author: Tel, 86-21-52230526; Fax, 86-21-62204010; E-mail, [email protected]Search for more papers by this authorThis work was supported by the grants from the Shanghai Rising-Star Program (No. 06QA14045) and the China National High-Tech Program (863) (No. 2006AA02Z115)
Abstract
Pyridoxal kinase is key enzyme for the biosynthesis of pyridoxa 15′-phosphate, the biologically active form of vitamin B6, in the salvage pathway. A pyridoxal kinase gene, BnPKL (GenBank accession No. DQ 463962), was isolated from oilseed rape (Brassica napus L.) following water stress through rapid amplification of complementary DNA (cDNA) ends. The results showed that the gene had two splice variants: PKL and PKL2.PKL, the long cDNA, encodes a 334 amino acid protein with a complete ATP-binding site, pyridoxal kinase-binding site and dimer interface site of a pyridoxal kinase, while PKL2, the short cDNA, lacked a partial domain. Southern blot showed that there were two copies in Brassica napus. The expression of BnPKL cDNA could rescue the mutant phenotype of Escherichia coli defective in pyridoxal kinase. Real-time reverse transcription-polymerase chain reaction revealed that the relative abundance of two transcripts are modulated by development and environmental stresses. Abscisic acid and NaCl were inclined to decrease PKL expression, but H2O2 and cold temperatures induced the PKL expression. In addition, the PKL expression could be transiently induced by jasmonate acid at an early stage, abscisic acid, salicylic acid and jasmonate acid enhanced the PKL expression in roots. Our results de monstrated that BnPKL was a pyridoxal kinase involved in responses to biotic and abiotic stresses.
References
- 1 John RA., Pyridoxal phosphate-dependent enzymes. Biochim Biophys Acta 1995, 1248: 81–96
- 2 Hill RE, Spenser ID. Biosynthesis of vitamin B. In: FC Neidhardt, RI Curtiss, JL Ingraham, ECC Lin, KB Low, B Magasanik, WS Reznikoff et al. eds. Washington : American Society for Microbiology, 1996.
- 3 Yang Y, Tsui HT, Man T, Winkler ME., Indentificaiton and function of the pkxY gene, which encodes a novel pyridoxal kinase involved in the salvage pathway of pyridoxal 5′-phosphate biosynthesis in Escherichia coli K-12. J Bacteriol 1998, 180: 1814–1812
- 4 Bean LE, Dvorachek WH, Braun EL, Errett A, Saenz GS, Giles MD, Werner-Washburne M et al., Analysis of the pdx-1 (snz-1/sno-1) region of the Neurospora crassa genome: correlation of pyridoxine-requiring phenotypes with mutations in two structural genes. Genetics 2001, 157: 1067–1075
- 5 Ehrenshaft M, Daub ME., Isolation of PDX2, a second novel gene in the pyridoxine biosynthesis pathway of eukaryotes, archaebacteria, and a subset of eubacteria. J Bacteriol 2001, 183: 3383–3390
- 6 Mittenhuber G., Phylogenetic analyses and comparative genomics of vitamin B6 (pyridoxine) and pyridoxal phosphate biosynthesis pathways. J Mol Microbiol Biotechnol 2001, 3: 1–20
- 7 Seack J, Perovic S, Gamulin V, Schroder HC, Beutelmann P, Muller IM, Muller WE., Identification of highly conserved genes: SNZ and SNO in the marine sponge Suberites domuncula: their gene structure and promoter activity in mammalian cells. Biochim Biophys Acta 2001, 1520: 21–34
- 8 Sivasubramaniam S, Vanniasingham VM, Tan CT, Chua NH., Characterisation of HEVER, a novel stress-induced gene from Hevea brasiliensis. Plant Mol Biol 1995, 29: 173–178
- 9 Denslow SA, Walls AA, Daub ME., Regulation of biosynthetic genes and antioxidant properties of vitamin B6 vitamers during plant defense responses. Physiol Mol Plant Pathol 2005, 66: 244–255
- 10 Denslow SA, Reuschhoff EE, Daub ME., Regulation of the Arabidopsis thaliana vitamin B6 biosynthesis genes by abiotic stress. Plant Physiol Biochem 2007, 45: 152–161
- 11 Tambasco-Studart M, Titiz O, Raschle T, Forster G, Amrhein N, Fitzpatrick TB., Vitamin B6 biosynthesis in higher plants. Proc Natl Acad Sci USA 2005, 102: 13687–13692
- 12 Gonzalez E, Danehower D, Daub M., Vitamer levels, stress response, enzyme activity, and gene regulation of Arabidopsis lines mutant in the pyridoxine/pridoxamine 5′-phosphate oxidase (PDX3) and the pyridoxal kinase (sos4) genes involved in the vitamin B6 salvage pathway. Plant Physiol 2007, 145: 985–996
- 13 Guirard BM, Snell EE., Physical and kinetic properties of pyridoxal reductase purified from bakers' yeast. Biofactors 1988, 1: 187–192
- 14 Morita T, Takegawa K, Yagi T., Disruption of the p1r1+ gene encoding pyridoxal reductase of Schizosaccharomyces pombe. J Biol Chem 2004, 135: 225–230
- 15 Drewke C, Leistner E. Biosynthesis of vitamin B6 and structurally related derivatives. In: G Litwack, T. Begley eds. Vitamins and Hormones, Advances in Research and Applications. San Diego : Academic Press, 2001.
- 16 Braun EL, Fuge EK, Padilla PA, Werner-Washburne M., A stationary-phase gene in Sacchromyces cerevisiae is a member of novel, highly conserved gene family. J Bacteriol 1996, 178: 6865–6872
- 17 Padilla PA, Fuge EK, Crawford ME, Errett A, Werner-Washburne M., The highly conserved, coregulated SNO and SNZ gene families in Saccharomyces cerevisiae respond to nutrient limitation. J Bacteriol 1998, 180: 5718–5726
- 18 Antelmann H, Bernhardt J, Schmid R, Mach H, Volder U, Hecker M., First step from a two-dimensional protein index towards a response-regulation map for Bacellus subtilis. Electrophoresis 1997, 18: 1451–1463
- 19 Chen D, Toone WM, Mata J, Lyne R, Burns G, Kivinen K., Global transcriptional responses of fission yeast to environmental stress. Mol Biol Cell 2003, 14: 214–229
- 20 Kannan K, Jain SK., Effect of vitamin B6 on oxygen radicals, mitochondrial membrane potential, and lipid peroxidation in H2O2-treated U937 monocytes. Free Radic Biol Med 2004, 36: 423–428
- 21 Lakshmi R, Lakshmi AV, Divan PV, Bamji MS., Effect of riboflavin or pyridoxine deficiency on inflammatory response. Indian J Biochem Biophys 1991, 28: 481–484
- 22 Chen H, Xiang L., Pyridoxine is required for post-embryonic root development and tolerance to osmotic and oxidative stress. Plant J 2005, 44: 396–408
- 23 Titiz O, Tambasco-Studart M, Warzych E, Apel K, Amrhein N, Laloi C, Fitzpatrick TB., PDX1 is essential for vitamin B6 biosynthesis, development and stress tolerance in Arabidopsis. Plant J 2006, 48: 933–946
- 24 Shi H, Xiong L, Stevenson B, Lu T, Zhu JK., The Arabidopsis salt overly sensitive 4 mutants uncover a critical role for vitamin B6 in plant salt tolerance. Plant Cell 2002, 14: 575–588
- 25 Shi H, Zhu JK., SOS4, a pyridoxal kinase gene, is required for root hair development in Arabidopsis. Plant Physiol 2002, 129: 585–593
- 26 Yang Y, Zhao G, Winkler ME., Identification of the pdxK gene that encodes pyridoxine (vitamin B6) kinase in Escherichia coli K-12. FEMS Microbiol Lett 1996, 141: 89–95
- 27 Guo K, Newell PC., Pyridoxal kinase knockout of Dictyostelium complemented by the human homologue. FEMS Microbiol Lett 2000, 189: 195–200
- 28 Kwok F, Churchich JE., Interaction between pyridoxal kinase and pyridoxine-5-P oxidase, two enzymes involved in the metabolism of vitamin B6. J Biol Chem 1980, 255: 882–887
- 29 Cash CD, Maitre M, Rumigny JF, Mandel P., Rapid purification by affinity chromatography of rat brain pyridoxal kinase and pyridoxamine-5-phosphate oxidase. Biochem Biophys Res Commun 1980, 96: 1755–1760
- 30 Hanna MC, Turner AJ, Kirkness EF., Human pyridoxal kinase. cDNA cloning, expression, and modulation by ligands of the benzodiazepine receptor. J Biol Chem 1997, 272: 10756–10760
- 31 Li MH, Kwok F, Chang WR, Lau CK, Zhang JP, Lo SCL, Jiang T et al., Crystal structure of brain pyridoxal kinase, a novel member of the ribokinase superfamily. J Biol Chem 2002, 277: 46385–46390
- 32 Lum HK, Kwok F, Lo SC., Cloning and characterization of Arabidopsis thaliana pyridoxal kinase. Planta 2002, 215: 870–879
- 33 Wang H, Liu D, Liu C, Zhang A., The pyridoxal kinase gene TaPdxK from wheat complements vitamin B6 synthesis-defective Escherichia coli. J Plant Physiol 2004, 161: 1053–1060
- 34 Ge T, Yu Y., Studies on nutritional requirements of regenerated plantlets in wheat. J Huazhong Agri Univ 1994, 13: 344–348
- 35 Jiang C, Chen Y., A method of isolation genomic DNA from Brassica. Chinese J Oil Crop Sci 1995, 17: 34–36
- 36 Livak KJ, Schmittgen TD., Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔ CT Method. Methods 2001, 25: 402–408
- 37 Altschul SF, Madden TL, Schäffer AA, Zhang J, Zhang Z, Miller W, Lipman DJ., Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res 1997, 25: 3389–3402
- 38 Schwede T, Kopp J, Guex N, Peitsch MC., SWISS-MODEL: an automated protein homology-modeling server. Nucleic Acids Res 2003, 31: 3381–3385
- 39 Yu S, Liu H, Luo L., Analysis of relative gene expression using different real-time quantitative PCR. Acta Agronom Sinica 2007, 33: 1214–1218
- 40 Schoenlein PV, Roa BB, Winkler ME., Divergent transcription of pdxB and homology between the pdxB and sera gene products in Escherichia coli K-12. J Bacteriol 1989, 171: 6084–6092
- 41 Lam HM, Winkler ME., Metabolic relationships between pyridoxine (vitamin B6) and serine biosynthesis in Escherichia coli K-12. J Bacteriol 1990, 172: 6518–6528
- 42 Turner JG, Ellis C, Devoto A., The jasmonate signal pathway. Plant Cell 2002, 14: S153–S164
