Russian Federation
An annotated gene of cytosolic NADP-dependent IDH of Hordeum vulgare was found in the NCBI database. This gene is localized in chromosome 8 and contains 15 exons and 14 introns. The sequence obtained after intron removal and exon fusion was used to select Real-time PCR primers. Optimization of primers showed that the optimal temperature for annealing is 60°C. A study of the promoter of this gene revealed the presence of various regulatory elements, such as a G-box, W-box, DRE, which indicates an important role of cIDH in plant adaptation to stress and possible light regulation of enzyme gene expression. Using the Methprimer program, a CpG island was found in the promoter, which suggests possible epigenetic regulation of expression through methylation/demethylation.
isocitrate dehydrogenase, promoter, cis-elements, primers, PCR
1. Galkin A. P. Regulyatornye oblasti promotorov genov rasteniy i belki —regulyatory promotornoy aktivnosti / A. P. Galkin // Biopolimery i kletka. – 2004. – T. 20. – №. 5. – S. 363.
2. Chen R. D. Chromatographic and immunological evidence that chloroplastic and cytosolic pea (Pisumsativum L.) NADP-isocitrate dehydrogenases are distinct isoenzymes / R.D. Chen et al. // Planta. – 1989. – V. 178. – P. 157-163.
3. Corpas F. J. NADPH-generating dehydrogenases: their role in the mechanism of protection against nitro-oxidative stress induced by adverse environmental conditions / F. J. Corpas, J.B. Barroso // Frontiers in Environmental Science. – 2014. – V. 2. – P. 55.
4. Corpas F. J. Peroxisomal plant metabolism–an update on nitric oxide, Ca2+ and the NADPH recycling network / F. J. Corpas, J. B. Barroso // Journal of Cell Science. – 2018. – V. 131. – №. 2. – P. jcs202978.
5. Hodges M. Higher plant NADP+-dependent isocitrate dehydrogenases, ammonium assimilation and NADPH production / M. Hodges et al. // Plant Physiology and Biochemistry. – 2003. – V. 41. – №. 6-7. – P. 577-585.
6. Igamberdiev A. U. Regulation of NAD-and NADP-dependent isocitrate dehydrogenases by reduction levels of pyridine nucleotides in mitochondria and cytosol of pea leaves / A.U. Igamberdiev, P. Gardeström // Biochimica et BiophysicaActa (BBA)-Bioenergetics.
7. Leterrier M. Cytosolic NADP-isocitrate dehydrogenase of pea plants: genomic clone characterization and functional analysis under abiotic stress conditions / M. Leterrier et al. // Free Radical Research. – 2007. – V. 41. – №. 2. – P. 191-199.
8. McKinnon J. D. Dual-Targeting of NADP+ - IsocitrateDehydrogenase : dis. – University of Saskatchewan, 2009.
9. Valderrama R. et al. The dehydrogenase‐mediated recycling of NADPH is a key antioxidant system against salt‐induced oxidative stress in olive plants / R. Valderrama et al. // Plant, Cell & Environment. – 2006. – V. 29. – №. 7. – P. 1449-1459
10. XuX. Structures of human cytosolic NADP-dependent isocitrate dehydrogenase reveal a novel self-regulatory mechanism of activity / X. Xu et al. // Journal of Biological Chemistry. – 2004. – V. 279, №. 32. – P. 33946-33957.
