GA3 mediated enhanced transcriptional rate and mRNA stability of 3-hydroxy-3-methylglutaryl coenzyme a reductase 1 (NtHMGR1) in Nicotiana tabacum L.

Authors

  • Raman MANOHARLAL ITC Limited, ITC Life Science and Technology Centre (LSTC), Peenya Industrial Area, 1st Phase, Bengaluru-560058, Karnataka
  • Lucky DUHAN Maharshi Dayanand University, Department of Biochemistry, Rohtak-124001
  • Ritu PASRIJA Maharshi Dayanand University, Department of Biochemistry, Rohtak-124001
  • Saiprasad V.S. GANDRA ITC Limited, ITC Life Science and Technology Centre (LSTC), Peenya Industrial Area, 1st Phase, Bengaluru-560058, Karnataka

DOI:

https://doi.org/10.55779/nsb14411317

Keywords:

GA3, HMGR1, Nicotiana tabacum, transcription run-on, transcriptional rate and mRNA stability

Abstract

Our present study evaluated the underlying molecular-mechanism(s) associated with the observed enhanced transcript levels and concomitant functional activity of 3-hydroxy-3-methylglutaryl coenzyme A reductase 1 (NtHMGR1), a rate-limiting enzyme of cytosolic mevalonate (MVA) pathway of terpenoids biosynthesis, by gibberellin A3 (GA3) treatment in model cultivated tobacco, Nicotiana tabacum L. Based on the transcription run-on and cordycepin chase assays, our results demonstrated that tobacco seeds-priming with GA3 causes a relative and significantly enhanced transcriptional rate and mRNA stability of NtHMGR1. Taken together, our study established that GA3 mediated transcriptional and post-transcriptional regulatory control as one of the mechanisms for the observed enhanced transcript-levels and consequently enhanced functional activity of NtHMGR1.

References

Abbas F, Ke Y, Yu R, Yue Y, Amanullah S, Jahangir MM, Fan Y (2017). Volatile terpenoids: multiple functions, biosynthesis, modulation and manipulation by genetic engineering. Planta 246:803-816. https://doi.org/10.1007/s00425-017-2749-x

Antolín-Llovera M, Leivar P, Arró M, Ferrer A, Boronat A, Campos N (2011). Modulation of plant HMG-CoA reductase by protein phosphatase 2A: positive and negative control at a key node of metabolism. Plant Signaling and Behavior 6:1127-1131. https://doi.org/10.4161/psb.6.8.16363

Aquil S, Husaini AM, Abdin MZ, Rather GM (2009). Overexpression of the HMG-CoA reductase gene leads to enhanced artemisinin biosynthesis in transgenic Artemisia annua plants. Planta Medica 75:1453-1458. https://doi.org/10.1055/s-0029-1185775

Atsumi G, Kagaya U, Tabayashi N, Matsumura T (2018). Analysis of the mechanisms regulating the expression of isoprenoid biosynthesis genes in hydroponically-grown Nicotiana benthamiana plants using virus-induced gene silencing. Scientific Reports 8:1-11. https://doi.org/10.1038/s41598-018-32901-5

Bach TJ (1986). Hydroxymethylglutaryl-CoA reductase, a key enzyme in phytosterol synthesis? Lipids 21:82-88. https://doi.org/10.1007/BF02534307

Bach TJ (1995). Some new aspects of isoprenoid biosynthesis in plants-a review. Lipids 30:191-202. https://doi.org/10.1007/BF02537822

Bansal S, Narnoliya LK, Mishra B, Chandra M, Yadav RK, Sangwan NS (2018). HMG-CoA reductase from Camphor Tulsi (Ocimum kilimandscharicum) regulated MVA dependent biosynthesis of diverse terpenoids in homologous and heterologous plant systems. Scientific Reports 8:1-15. https://doi.org/10.1038/s41598-017-17153-z

Brahmkshatriya PP, Brahmkshatriya PS (2013). Terpenes: Chemistry, biological role, and therapeutic applications. In: Natural Products: Phytochemistry, Botany and Metabolism of Alkaloids, Phenolics and Terpenes. Springer (Berlin Heidelberg): Berlin, Germany. https://doi.org/10.1007/978-3-642-22144-6_120

Brooker JD, Russell DW (1979). Regulation of microsomal 3-hydroxy-3-methylglutaryl coenzyme A reductase from pea seedlings: rapid posttranslational phytochrome-mediated decrease in activity and in vivo regulation by isoprenoid products. Archives of Biochemistry and Biophysics 198:323-334. https://doi.org/10.1016/0003-9861(79)90425-9

Campos N, Boronat A (1995). Targeting and topology in the membrane of plant 3-hydroxy-3-methylglutaryl coenzyme A reductase. The Plant Cell 7:2163-2174. https://doi.org/10.1105/tpc.7.12.2163

Chappell J, Wolf F, Proulx J, Cuellar R, Saunders C (1995). Is the reaction catalyzed by 3-hydroxy-3-methylglutaryl coenzyme A reductase a rate-limiting step for isoprenoid biosynthesis in plants? Plant Physiology 109:1337-1343. https://doi.org/10.1104/pp.109.4.1337

da Costa RF, Freire VN, Bezerra EM, Cavada BS, Caetano EW, de Lima Filho JL, Albuquerque EL (2012). Explaining statin inhibition effectiveness of HMG-CoA reductase by quantum biochemistry computations. Physical Chemistry and Chemical Physics 14:1389-1398. https://doi.org/10.1039/C1CP22824B

Dai Z, Cui G, Zhou S-F, Zhang X, Huang L (2011). Cloning and characterization of a novel 3-hydroxy-3-methylglutaryl coenzyme A reductase gene from Salvia miltiorrhiza involved in diterpenoid tanshinone accumulation. Journal of Plant Physiology 168:148-157. https://doi.org/10.1016/j.jplph.2010.06.008

Dale S, Arró M, Becerra B, Morrice NG, Boronat A, Hardie DG, Ferrer A (1995). Bacterial expression of the catalytic domain of 3-hydroxy-3-methylglutaryl-CoA reductase (isoform HMGR1) from Arabidopsis thaliana, and its inactivation by phosphorylation at Ser577 by Brassica oleracea 3-hydroxy-3-methylglutaryl-CoA reductase kinase. European Journal of Biochemistry 233:506-513. https://doi.org/10.1111/j.1432-1033.1995.506_2.x

Depuydt S, Hardtke CS (2011). Hormone signalling crosstalk in plant growth regulation. Current Biology 21:R365-R373. https://doi.org/10.1016/j.cub.2011.03.013

Diarra ST, He J, Wang J, Li J (2013). Ethylene treatment improves diosgenin accumulation in in vitro cultures of Dioscorea zingiberensis via up-regulation of CAS and HMGR gene expression. Electronic Journal of Biotechnology 16:6-6. https://doi.org/10.2225/vol16-issue5-fulltext-9

Espenshade PJ, Hughes AL (2007). Regulation of sterol synthesis in eukaryotes. Annual Review of Genetics 41:401-427. https://doi.org/10.1146/annurev.genet.41.110306.130315

Friesen JA, Rodwell VW (2004). The 3-hydroxy-3-methylglutaryl coenzyme-A (HMG-CoA) reductases. Genome Biology 5:248. https://doi.org/10.1186/gb-2004-5-11-248

Goldstein JL, Brown MS (1990). Regulation of the mevalonate pathway. Nature 343:425-430. https://doi.org/10.1038/343425a0

Gubler F, Chandler PM, White RG, Llewellyn DJ, Jacobsen JV (2002). Gibberellin signaling in barley aleurone cells. Control of SLN1 and GAMYB expression. Plant Physiology 129:191-200. https://doi.org/10.1104/pp.010918

Hemmerlin A (2013). Post-translational events and modifications regulating plant enzymes involved in isoprenoid precursor biosynthesis. Plant Science 203:41-54. https://doi.org/10.1016/j.plantsci.2012.12.008

Hemmerlin A, Bach TJ (1998). Effects of mevinolin on cell cycle progression and viability of tobacco BY-2 cells. The Plant Journal 14:65-74. https://doi.org/10.1046/j.1365-313X.1998.00095.x

Hemmerlin A, Bach TJ (2000). Farnesol-induced cell death and stimulation of 3-hydroxy-3-methylglutaryl-coenzyme A reductase activity in tobacco cv bright yellow-2 cells. Plant Physiology 123:1257-1268. https://doi.org/10.1104/pp.123.4.1257

Hemmerlin A, Gerber E, Feldtrauer JF, Wentzinger L, Hartmann MA, Tritsch D, ... Bach TJ (2004). A review of tobacco BY‐2 cells as an excellent system to study the synthesis and function of sterols and other isoprenoids. Lipids 39(8):723-735. https://doi.org/10.1007/s11745-004-1289-0

Hemmerlin A, Hoeffler JF, Meyer O, Tritsch D, Kagan IA, Grosdemange-Billiard C, ... Bach TJ (2003). Cross-talk between the cytosolic mevalonate and the plastidial methylerythritol phosphate pathways in tobacco bright yellow-2 cells. Journal of Biological Chemistry 278(29):26666-26676. https://doi.org/10.1074/jbc.M302526200

Holmberg N, Harker M, Gibbard CL, Wallace AD, Clayton JC, Rawlins S, ... Safford R (2002). Sterol C-24 methyltransferase type 1 controls the flux of carbon into sterol biosynthesis in tobacco seed. Plant Physiology 130(1):303-311. https://doi.org/10.1104/pp.004226

Jelesko JG, Jenkins SM, Rodríguez-Concepción M, Gruissem W (1999). Regulation of tomato HMG1 during cell proliferation and growth. Planta 208:310-318. https://doi.org/10.1007/s004250050564

Khan AA, Agarwal H, Reddy SS, Arige V, Natarajan B, Gupta V, ... Mahapatra NR (2020). MicroRNA 27a is a key modulator of cholesterol biosynthesis. Molecular and Cellular Biology 40(9):e00470-19. https://doi.org/10.1128/MCB.00470-19

Khan AA, Gupta V, Ananthamohan K, Arige V, Reddy SS, Barthwal MK, ... Mahapatra NR (2018). Post-transcriptional regulation of 3-hydroxy-3-methylglutaryl-coenzyme A reductase: crucial role of microRNA-27a or Identification of microRNA-27a as a key regulator of cholesterol homeostasis. bioRxiv 383448. https://doi.org/10.1101/383448

Kobayashi K, Suzuki M, Tang J, Nagata N, Ohyama K, Seki H, ... Muranaka T (2007). Lovastatin insensitive 1, a novel pentatricopeptide repeat protein, is a potential regulatory factor of isoprenoid biosynthesis in Arabidopsis. Plant and Cell Physiology 48(2):322-331. https://doi.org/10.1093/pcp/pcm005

Korth KL, Jaggard DA, Dixon RA (2000). Developmental and light-regulated post-translational control of 3-hydroxy-3-methylglutaryl-CoA reductase levels in potato. The Plant Journal 23:507-516. https://doi.org/10.1046/j.1365-313x.2000.00821.x

Kuzuyama T, Seto H (2012). Two distinct pathways for essential metabolic precursors for isoprenoid biosynthesis. Proceedings of the Japan Academy, Series B 88:41-52. https://doi.org/10.2183/pjab.88.41

Learned RM, Connolly EL (1997). Light modulates the spatial patterns of 3-hydroxy-3-methylglutaryl coenzyme A reductase gene expression in Arabidopsis thaliana. The Plant Journal 11:499-511. https://doi.org/10.1046/j.1365-313X.1997.11030499.x

Leivar P, González VM, Castel S, Trelease RN, López-Iglesias C, Arró M, ... Fernandez-Busquets X (2005). Subcellular localization of Arabidopsis 3-hydroxy-3-methylglutaryl-coenzyme A reductase. Plant Physiology 137(1):57-69. https://doi.org/10.1104/pp.104.050245

Leubner-Metzger G (2001). Brassinosteroids and gibberellins promote tobacco seed germination by distinct pathways. Planta 213:758-763. https://doi.org/10.1007/s004250100542

Li Z, Gao Y, Zhang Y, Lin C, Gong D, Guan Y, Hu J (2018). Reactive oxygen species and gibberellin acid mutual induction to regulate tobacco seed germination. Frontiers in Plant Science 9:1279. https://doi.org/10.3389/fpls.2018.01279

Liao P, Chen X, Wang M, Bach TJ, Chye M-L (2018). Improved fruit α-tocopherol, carotenoid, squalene and phytosterol contents through manipulation of Brassica juncea 3-HYDROXY-3-METHYLGLUTARYL-COA SYNTHASE 1 in transgenic tomato. Plant Biotechnology Journal 16:784-796. https://doi.org/10.1111/pbi.12828

Liao P, Wang H, Wang M, Hsiao A-S, Bach TJ, Chye M-L (2014). Transgenic tobacco overexpressing Brassica juncea HMG-CoA synthase 1 shows increased plant growth, pod size and seed yield. PLoS One 9:e98264. https://doi.org/10.1371/journal.pone.0098264

Lichtenthaler HK, Rohmer M, Schwender J (1997). Two independent biochemical pathways for isopentenyl diphosphate and isoprenoid biosynthesis in higher plants. Physiologia Plantarum 101:643-652. https://doi.org/10.1111/j.1399-3054.1997.tb01049.x

Loguercio LL, Scott HC, Trolinder NL, Wilkins TA (1999). HMG-CoA reductase gene family in cotton (Gossypium hirsutum L.): unique structural features and differential expression of hmg2 potentially associated with synthesis of specific isoprenoids in developing embryos. Plant and Cell Physiology 40:750-761. https://doi.org/10.1093/oxfordjournals.pcp.a029602

Manoharlal R, Saiprasad G (2019). Assessment of germination, phytochemicals, and transcriptional responses to ethephon priming in soybean [Glycine max (L.) Merrill]. Genome 62:769-783. https://doi.org/10.1139/gen-2019-0013

Manoharlal R, Saiprasad G, Kaikala V (2019). Gibberellin A3 mediated decreased transcriptional rate, mRNA stability and non-competitive inhibition of DNA methyltransferases in tobacco. Biologia Plantarum 63:343-353. https://doi.org/10.32615/bp.2019.040

Manoharlal R, Saiprasad G, Kaikala V, Kumar RS, Kovařík A (2018a). Analysis of DNA methylome and transcriptome profiling following Gibberellin A3 (GA3) foliar application in Nicotiana tabacum L. Indian Journal of Plant Physiology 23:543-556. https://doi.org/10.1007/s40502-018-0393-5

Manoharlal R, Saiprasad G, Thambrahalli A, Madhavakrishna K (2018b). Dissecting the transcriptional networks underlying the gibberellin response in Nicotiana tabacum. Biologia Plantarum 62:647-662. https://doi.org/10.1007/s10535-018-0809-0

Manoharlal R, Saiprasad G, Ullagaddi C, Kovařík A (2018c). Gibberellin A3 as an epigenetic determinant of global DNA hypo-methylation in tobacco. Biologia Plantarum 62:11-23. https://doi.org/10.1007/s10535-017-0738-3

Mansouri H, Asrar Z, Amarowicz R (2011). The response of terpenoids to exogenous gibberellic acid in Cannabis sativa L. at vegetative stage. Acta Physiologiae Plantarum 33:1085-1091. https://doi.org/10.1007/s11738-010-0636-1

Merret R, Cirioni J-R, Bach TJ, Hemmerlin A (2007). A serine involved in actin-dependent subcellular localization of a stress-induced tobacco BY-2 hydroxymethylglutaryl-CoA reductase isoform. FEBS Letters 581:5295-5299. https://doi.org/10.1016/j.febslet.2007.10.023

Ness GC (2015). Physiological feedback regulation of cholesterol biosynthesis: Role of translational control of hepatic HMG-CoA reductase and possible involvement of oxylanosterols. Biochimica et Biophysica Acta (BBA)-Molecular and Cell Biology of Lipids 1851:667-673. https://doi.org/10.1016/j.bbalip.2015.02.008

Nieto B, Forés O, Arró M, Ferrer A (2009). Arabidopsis 3-hydroxy-3-methylglutaryl-CoA reductase is regulated at the post-translational level in response to alterations of the sphingolipid and the sterol biosynthetic pathways. Phytochemistry 70:53-59. https://doi.org/10.1016/j.phytochem.2008.10.010

Pagare S, Bhatia M, Tripathi N, Pagare S, Bansal Y (2015). Secondary metabolites of plants and their role: Overview. Current Trends in Biotechnology and Pharmacy 9:293-304.

Pichersky E, Raguso RA (2018). Why do plants produce so many terpenoid compounds? New Phytologist 220:692-702. https://doi.org/10.1111/nph.14178

Pu G-B et al. (2009). Salicylic acid activates artemisinin biosynthesis in Artemisia annua L. Plant Cell Reports 28:1127-1135. https://doi.org/10.1007/s00299-009-0713-3

Russell D, Knight J, Wilson T (1985). Pea seedling HMG-CoA reductases: regulation of activity in vitro by phosphorylation and Ca2+ and postranslational control in vivo by phytochrome and isoprenoid hormones. Current topics in plant biochemistry and physiology: Proceedings of the Plant Biochemistry and Physiology Symposium held at the University of Missouri, Columbia.

Samad AFA, Rahnamaie-Tajadod R, Sajad M, Jani J, Murad AMA, Noor NM, Ismail I (2019). Regulation of terpenoid biosynthesis by miRNA in Persicaria minor induced by Fusarium oxysporum. BMC Genomics 20:586. https://doi.org/10.1186/s12864-019-5954-0

Shaikh S, Shriram V, Khare T, Kumar V (2020). Biotic elicitors enhance diosgenin production in Helicteres isora L. suspension cultures via up-regulation of CAS and HMGR genes. Physiology and Molecular Biology of Plants 26:593-604. https://doi.org/10.1007/s12298-020-00774-6

Shi L, Olszewski NE (1998). Gibberellin and abscisic acid regulate GAST1 expression at the level of transcription. Plant Molecular Biology 38:1053-1060. https://doi.org/10.1023/A:1006007315718

Skoog F, Miller C (1957). Chemical regulation of growth and organ formation in plant tissues cultured in vitro. In: Symp Soc Exp Biol., pp 118-131

Soma Y, Tsuruno K, Wada M, Yokota A, Hanai T (2014). Metabolic flux redirection from a central metabolic pathway toward a synthetic pathway using a metabolic toggle switch. Metabolic Engineering 23:175-184. https://doi.org/10.1016/j.ymben.2014.02.008

Stermer BA, Bianchini GM, Korth KL (1994). Regulation of HMG-CoA reductase activity in plants. Journal of Lipid Research 35:1133-1140. https://doi.org/10.1016/S0022-2275(20)39958-2

Su S et al. (2018). Feedback regulation of 3-Hydroxy-3-methylglutaryl-coenzyme A reductase in livers of mice. Arteriosclerosis, Thrombosis, and Vascular Biology 38:A457-A457. https://doi.org/10.1161/atvb.38.suppl_1.457

Suzuki M et al. (2009). Complete blockage of the mevalonate pathway results in male gametophyte lethality. Journal of Experimental Botany 60:2055-2064. https://doi.org/10.1093/jxb/erp073

Tang Y, Zhong L, Wang X, Zheng H, Chen L (2019). Molecular identification and expression of sesquiterpene pathway genes responsible for patchoulol biosynthesis and regulation in Pogostemon cablin. Botanical Studies 60:1-11. https://doi.org/10.1186/s40529-019-0259-9

Tholl D (2015) Biosynthesis and biological functions of terpenoids in plants. In: Biotechnology of isoprenoids. Springer, pp 63-106 https://doi.org/10.1007/10_2014_295

Tohge T, Scossa F, Fernie AR (2015). Integrative approaches to enhance understanding of plant metabolic pathway structure and regulation. Plant Physiology 169:1499-1511. https://doi.org/10.1104/pp.15.01006

Vranová E, Coman D, Gruissem W (2013). Network analysis of the MVA and MEP pathways for isoprenoid synthesis. Annual Review of Plant Biology 64:665-700. https://doi.org/10.1146/annurev-arplant-050312-120116

Wagner K-H, Elmadfa I (2003). Biological relevance of terpenoids. Annals of Nutrition and Metabolism 47:95-106. https://doi.org/10.1159/000070030

Wang Y, Guo B, Zhang F, Yao H, Miao Z, Tang K (2007). Molecular cloning and functional analysis of the gene encoding 3-hydroxy-3-methylglutaryl coenzyme A reductase from hazel (Corylus avellana L. Gasaway). Journal of Biochemistry and Molecular Biology 40:861. https://doi.org/10.5483/BMBRep.2007.40.6.861

Wei H, Xu C, Movahedi A, Sun W, Li D, Zhuge Q (2019). Characterization and function of Hydroxy-3-Methylglutaryl-CoA Reductase in Populus trichocarpa: Overexpression of PtHMGR enhances terpenoids in transgenic poplar. Frontiers in Plant Science 10:1476. https://doi.org/10.3389/fpls.2019.01476

Wei J, Yang J, Ling M, Liu H, Zhan R, Chen W (2013). Regulatory effect of methyl jasmonate on HMGR, DXR and DXS genes expression in Amomum villosum Lour. Journal of Guangzhou University of Traditional Chinese Medicine 30:88-92.

Wentzinger LF, Bach TJ, Hartmann M-A (2002). Inhibition of squalene synthase and squalene epoxidase in tobacco cells triggers an up-regulation of 3-hydroxy-3-methylglutaryl coenzyme A reductase. Plant Physiology 130:334-346. https://doi.org/10.1104/pp.004655

Wink M (2010). Introduction: biochemistry, physiology and ecological functions of secondary metabolites. Annual Plant Reviews: Biochemistry of Plant Secondary Metabolism 40:1-19.https://doi.org/10.1002/9781444320503

Wong RJ, Mc Cormack DK, Russell DW (1982). Plastid 3-hydroxy-3-methylglutaryl coenzyme A reductase has distinctive kinetic and regulatory features: properties of the enzyme and positive phytochrome control of activity in pea seedlings. Archives of Biochemistry and Biophysics 216:631-638. https://doi.org/10.1016/0003-9861(82)90253-3

Zhang H, Wang H, Zhu Q, Gao Y, Wang H, Zhao L, ... Gu L (2018). Transcriptome characterization of moso bamboo (Phyllostachys edulis) seedlings in response to exogenous gibberellin applications. BMC Plant Biology 18(1):1-15. https://doi.org/10.1186/s12870-018-1336-z

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Published

2022-11-28

How to Cite

MANOHARLAL, R., DUHAN, L., PASRIJA, R., & GANDRA, S. V. . (2022). GA3 mediated enhanced transcriptional rate and mRNA stability of 3-hydroxy-3-methylglutaryl coenzyme a reductase 1 (NtHMGR1) in Nicotiana tabacum L. Notulae Scientia Biologicae, 14(4), 11317. https://doi.org/10.55779/nsb14411317

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