BMB Reports

생화학분자생물학회 (Korean Society for Biochemistry and Molecular Biology)
권호 표지
DOI QR코드

DOI QR Code

Molecular Cloning and Functional Analysis of the Gene Encoding 3-hydroxy-3-methylglutaryl Coenzyme A Reductase from Hazel (Corylus avellana L. Gasaway)

  • Wang, Yechun (Plant Biotechnology Research Center, School of Agriculture and Biology, School of Life Science and Technology, Fudan-SJTU-Nottingham Plant Biotechnology R&D Center, Shanghai Jiao Tong University) ;
  • Guo, Binhui (Plant Biotechnology Research Center, School of Agriculture and Biology, School of Life Science and Technology, Fudan-SJTU-Nottingham Plant Biotechnology R&D Center, Shanghai Jiao Tong University) ;
  • Zhang, Fei (Plant Biotechnology Research Center, School of Agriculture and Biology, School of Life Science and Technology, Fudan-SJTU-Nottingham Plant Biotechnology R&D Center, Shanghai Jiao Tong University) ;
  • Yao, Hongyan (Plant Biotechnology Research Center, School of Agriculture and Biology, School of Life Science and Technology, Fudan-SJTU-Nottingham Plant Biotechnology R&D Center, Shanghai Jiao Tong University) ;
  • Miao, Zhiqi (Plant Biotechnology Research Center, School of Agriculture and Biology, School of Life Science and Technology, Fudan-SJTU-Nottingham Plant Biotechnology R&D Center, Shanghai Jiao Tong University) ;
  • Tang, Kexuan (Plant Biotechnology Research Center, School of Agriculture and Biology, School of Life Science and Technology, Fudan-SJTU-Nottingham Plant Biotechnology R&D Center, Shanghai Jiao Tong University)
  • 발행 : 2007.11.30
PDF 다운로드
⟨ 이전 논문 다음 논문 ⟩

초록

The enzyme 3-hydroxy-3-methylglutaryl-CoA reductase (HMGR; EC1.1.1.34) catalyzes the first committed step of isoprenoids biosynthesis in MVA pathway. Here we report for the first time the cloning and characterization of a full-length cDNA encoding HMGR (designated as CgHMGR, GenBank accession number EF206343) from hazel (Corylus avellana L. Gasaway), a taxol-producing plant species. The full-length cDNA of CgHMGR was 2064 bp containing a 1704-bp ORF encoding 567 amino acids. Bioinformatic analyses revealed that the deduced CgHMGR had extensive homology with other plant HMGRs and contained two transmembrane domains and a catalytic domain. The predicted 3-D model of CgHMGR had a typical spatial structure of HMGRs. Southern blot analysis indicated that CgHMGR belonged to a small gene family. Expression analysis revealed that CgHMGR expressed high in roots, and low in leaves and stems, and the expression of CgHMGR could be up-regulated by methyl jasmonate (MeJA). The functional color assay in Escherichia coli showed that CgHMGR could accelerate the biosynthesis of $\beta$-carotene, indicating that CgHMGR encoded a functional protein. The cloning, characterization and functional analysis of CgHMGR gene will enable us to further understand the role of CgHMGR involved in taxol biosynthetic pathway in C. avellana at molecular level.

키워드

참고문헌

  1. Aoyagi, K., Beyou, A., Moon, K., Fang, L. and Ulrich, T. (1993) Isolation and characterization of cDNAs encoding wheat 3-hydroxy-3-methylglutaryl coenzyme A reductase. Plant Physiol. 102, 623-628. https://doi.org/10.1104/pp.102.2.623
  2. Baebler, S., Camloh, M., Kovac, M., Ravnikar, M. and Zel, J. (2002) Jasmonic acid stimulates taxane production in cell suspension culture of yew (Taxus x media). Planta Med. 68, 475-476. https://doi.org/10.1055/s-2002-32083
  3. Boone, R. B., Hoffman, A., Daley, L. and Arbogast, B. (2000) How Taxol was found in hazelnut trees. Australian Nutgrower. 14, 36.
  4. Bestoso, F., Ottaggio, L., Armirotti, A., Balbi, A., Damonte, G., Degan, P., Mazzei, M., Cavalli, F., Ledda, B. and Miele, M. (2006) In vitro cell cultures obtained from different explants of Corylus avellana produce Taxol and taxanes. BMC Biotechnol. 6, 45. doi:10.1186/1472-6750-6-45.
  5. Bochar, D. A., Stauffacher, C. V. and Rodwell, V. W. (1999) Sequence comparisons reveal two classes of 3-hydroxy-3-methylglutaryl coenzyme A reductase. Mol. Genet. Metab. 66, 122-127. https://doi.org/10.1006/mgme.1998.2786
  6. Chappell, J. (1995) The biochemistry and molecular biology of isoprenoid metabolism. Plant Physiol. 107, 1-6. https://doi.org/10.1104/pp.107.1.1
  7. Chye, M. L., Tan, C. T. and Chua, N. H. (1992) Three genes encode 3-hydroxy -3-methylglutaryl coenzyme A reductase in Hevea brasiliensis: hmg1 and hmg3 are differentially expressed. Plant Mol. Biol. 19, 473-484. https://doi.org/10.1007/BF00023395
  8. Denbow, C. J., Lang, S. and Cramer, C. L. (1996) The N-terminal domain of tomato 3-hydroxy-3-methylglutaryl coenzyme A reductase. J. Biol. Chem. 271, 9710-9715. https://doi.org/10.1074/jbc.271.16.9710
  9. Enjuto, M., Balcells, L., Campos, N., Caelles, C., Arro, M. and Boronat, A. (1994) Arabidopsis thaliana contains two differentially expressed 3-hydroxy-3-methylglutaryl-CoA reductase genes, which encode microsomal forms of the enzyme. Proc. Natl. Acad. Sci. USA 91, 927-931. https://doi.org/10.1073/pnas.91.3.927
  10. Eisenreich, W., Menhard, B., Hylands, P. J., Zenk, M. H. and Bacher, A. (1996) Studies on the biosynthesis of taxol: the taxane carbon skeleton is not of mevalonoid origin. Proc. Natl. Acad. Sci. USA 93, 6431-6436. https://doi.org/10.1073/pnas.93.13.6431
  11. Eisenreich, W., Schwarz, M., Cartayrade, A., Arigoni, D., Zenk, M. H. and Bacher, A. (1998) The deoxyxylulose phosphate pathway of terpenoid biosynthesis in plants and microorganisms. Chem. Biol. 5, 221-R233. https://doi.org/10.1016/S1074-5521(98)90002-3
  12. Hoffman, A., Khan, W., Worapong, J., Strobel, G., Griffin, D., Arbogast, B., Barofsky, D., Boone, R. B., Ning, L., Zheng, P. and Daley, L. (1998) Bioprospecting for Taxol in angiosperm plant extracts. Spectroscopy 13, 22-32.
  13. Istvan, E. S., Palnitkar, M., Buchanan, S. K. and Deisenhofer, J. (2000) Crystal structure of the catalytic portion of human HMGCoA reductase: insights into regulation of activity and catalysis. EMBO J. 19, 819-830. https://doi.org/10.1093/emboj/19.5.819
  14. Jiang, J. H., Kai, G. Y., Cao, X. Y., Chen, F. M., He, D. N. and Liu, Q. (2006) Molecular cloning of a HMG-CoA reductase gene from Eucommia ulmoides Oliver. Biosci Rep. 26, 171-181. https://doi.org/10.1007/s10540-006-9010-3
  15. Jaakola, L., Pirttila, A. M., Halonen, M. and Hohtola, A. (2001) Isolation of high quality RNA from bilberry (Vaccinium myrtillus L.) fruit. Mol. Biotechnol. 19, 201-203. https://doi.org/10.1385/MB:19:2:201
  16. Jain, A. K., Vincent, R. M. and Nessler, C. L. (2000) Molecular characterization of a hydroxymethylglutaryl-CoA reductase gene from mulberry (Morus alba L.). Plant Mol. Biol. 42, 559-569. https://doi.org/10.1023/A:1006336825226
  17. Jennewein, S., Wildung, M. R., Chau, M., Walker, K. and Croteau, R. (2004) Random sequencing of an induced Taxus cell cDNA library for identification of clones involved in Taxol biosynthesis. Proc. Natl. Acad. Sci. USA 101, 9149-9154. https://doi.org/10.1073/pnas.0403009101
  18. Kato-Emori, S., Higashi, K., Hosoya, K., Kobayashi, T. and Ezura, H. (2001) Cloning and characterization of the gene encoding 3-hydroxy-3-methylglutaryl coenzyme A reductase in melon (Cucumis melo L. reticulatus). Mol. Genet. Genomics 265, 135-142. https://doi.org/10.1007/s004380000401
  19. Korth, K. L., Stemer, B. A., Bhattacharyya, M. K. and Dixon, R. A. (1997) HMG-CoA reductase gene families that differentially accumulate transcripts in potato tubers are developmentally expressed in floral tissues. Plant Mol. Biol. 33, 545-551. https://doi.org/10.1023/A:1005743011651
  20. Laule, O., Furholz, A., Chang, H. S., Zhu, T., Wang, X., Heifetz, P. B., Gruissem, W. and Lange, B. M. (2003) Crosstalk between cytosolic and plastidial pathways of isoprenoid biosynthesis in Arabidopsis thaliana. Proc. Natl. Acad. Sci. USA 100, 6866-6871. https://doi.org/10.1073/pnas.1031755100
  21. Lansing, A., Haertel, M., Gordon, M. and Floss, H. G. (1991) Biosynthetic studies on taxol. Planta Med. 57, 83. https://doi.org/10.1055/s-2006-960028
  22. Liao, Z. H., Tan, Q. M., Chai, Y. R., Zuo, K. J., Chen, M., Gong, Y. F, Wang, P., Pi, Y., Tang, F., Sun, X. F. and Tang, K. X. (2004) Cloning and chracterisation of the gene encoding HMG-CoA reductase from Taxus media and its functional identification in yeast. Funct. Plant Biol. 31, 73-81. https://doi.org/10.1071/FP03153
  23. Liu, G. M., Fang, W. S., Qian, J. F. and Lan, H. (2001) Distribution of paclitaxel and its congeners in Taxus mairei. Fitoterapia 72, 743-746. https://doi.org/10.1016/S0367-326X(01)00302-1
  24. Murashige, T. and Skoog, F. (1962) A revised medium for rapid growth and bioassay with tobacco tissue cultures. Physiol. Plant. 15, 473-497. https://doi.org/10.1111/j.1399-3054.1962.tb08052.x
  25. Matthews, P. D. and Wurtzel, E. T. (2000) Metabolic engineering of carotenoid accumulation in Escherichia coli by modulation of the isoprenoid precursor pool with expression of deoxyxylulose phosphate synthase. Appl. Microbiol. Biotechnol. 53, 396-400. https://doi.org/10.1007/s002530051632
  26. Misawa, N., Satomi, Y., Kondo, K., Yokoyama, A., Kajiwara, S., Saito, T., Ohtani, T. and Miki, W. (1995) Structure and functional analysis of a marine bacterial carotenoid biosynthesis gene cluster and astaxanthin biosynthetic pathway proposed at the gene level. J. Bacteriol. 177, 6575-6584. https://doi.org/10.1128/jb.177.22.6575-6584.1995
  27. Palazon, J., Cusido, R. M., Bonfill, M., Morales, C. and Pinol, M. T. (2003) Inhibition of paclitaxel and baccatin III accumulation by mevinolin and fosmidomycin in suspension cultures of Taxus baccata. J. Biotechnol. 101, 157-163. https://doi.org/10.1016/S0168-1656(02)00318-8
  28. Paterson, A. H., Curt, L. B. and Wendel, J. F. (1993) A rapid method for extraction of cotton (Gossypium ssp.) genomic DNA suitable for RFLP and PCR analysis. Plant Mol. Biol. Rep. 11, 122-127. https://doi.org/10.1007/BF02670470
  29. Rohmer, M., Knani, M., Simonin, P., Sutter, B. and Sahm, H. (1993) Isoprenoid biosynthesis in bacteria: a novel pathway for the early steps leading to isopentenyl diphosphate. Biochem. J. 295, 517-524. https://doi.org/10.1042/bj2950517
  30. Ro, D. K., Paradise, E. M., Ouellet, M., Fisher, K. J., Newman, K. L., Ndungu, J. M., Ho, K. A., Eachus, R. A., Ham, T. S., Kirby, J., Chang, M. C., Withers, S. T., Shiba, Y., Sarpong, R. and Keasling, J. D. (2006) Production of the antimalarial drug precursor artemisinic acid in engineered yeast. Nature 440, 940-943. https://doi.org/10.1038/nature04640
  31. Service, R. F. (2000) Hazel trees offer new source of cancer drug. Science 288, 27-28.
  32. Schwede, T., Kopp, J., Guex, N. and Peitsch, M. C. (2003) SWISSMODEL: an automated protein homology-modeling server. Nucleic Acids Res. 31, 3381-3385. https://doi.org/10.1093/nar/gkg520
  33. Shen, G. A., Pang, Y. Z., Wu, W. S., Liao, Z. H., Zhao, L. X., Sun, X. F. and Tang, K. X. (2006) Cloning and characterization of a rootspecific expressing gene encoding 3-hydroxy-3-methylglutaryl coenzyme a reductase from Ginkgo biloba. Mol. Biol. Rep. 33, 117-127. https://doi.org/10.1007/s11033-006-0014-7
  34. Wani, M. C., Taylor, H. L., Wall, M. E., Coggon, P. and McPhail, A. (1971) Plant antitumor agents. VI. The isolation and structure of Taxol, a novel antileukemic and antitumor agent from Taxus brevifolia. J. Am. Chem. Soc. 93, 2325-2327. https://doi.org/10.1021/ja00738a045
  35. Yukimune, Y., Tabata, H., Higashi, Y. and Hara, Y. (1996) Methyl jasmonate induced overproduction of paclitaxel and baccatin III in Taxus cell suspension cultures. Nat. Biotechnol. 14, 1129-1132. https://doi.org/10.1038/nbt0996-1129
  36. Zamir, L. O., Nedea, M. E. and Garneau, F. X. (1992) Biosynthetic building blocks of Taxus canadensis taxanes. Tetrahedron Lett. 33, 5235-5236. https://doi.org/10.1016/S0040-4039(00)79141-6

피인용 문헌

  1. Characterization and Expression of Two cDNA Encoding 3-Hydroxy-3-methylglutaryl coenzyme A Reductase Isoforms in Coffee (Coffea arabicaL.) vol.15, pp.10, 2011, https://doi.org/10.1089/omi.2010.0140
  2. Expression analysis of steroid pathway genes revealed positive correlation with diosgenin biosynthesis in Trillium govanianum vol.38, pp.11, 2016, https://doi.org/10.1007/s11738-016-2289-1
  3. Cloning and functional characterization of 3-hydroxy-3-methylglutaryl coenzyme A reductase gene from Withania somnifera: an important medicinal plant vol.250, pp.2, 2013, https://doi.org/10.1007/s00709-012-0450-2
  4. Characterization and functional analysis of the genes encoding 1-deoxy-d-xylulose-5-phosphate reductoisomerase and 1-deoxy-d-xylulose-5-phosphate synthase, the two enzymes in the MEP pathway, from Amomum villosum Lour vol.39, pp.8, 2012, https://doi.org/10.1007/s11033-012-1676-y
  5. Light and temperature regulated terpene biosynthesis: hepatoprotective monoterpene picroside accumulation in Picrorhiza kurrooa vol.10, pp.3, 2010, https://doi.org/10.1007/s10142-009-0152-9
  6. Molecular cloning, characterization and function analysis of the gene encoding HMG-CoA reductase from Euphorbia Pekinensis Rupr vol.37, pp.3, 2010, https://doi.org/10.1007/s11033-009-9558-7
  7. Phytochemistry and biosynthesis of δ-lactone withanolides vol.15, pp.5, 2016, https://doi.org/10.1007/s11101-015-9420-6
  8. Validation and application of reference genes for quantitative gene expression analyses in various tissues of Bupleurum chinense vol.38, pp.8, 2011, https://doi.org/10.1007/s11033-010-0648-3
  9. Molecular cloning, characterization and expression analysis of 3-hydroxy-3-methylglutaryl coenzyme A reductase gene from Centella asiatica L. vol.42, pp.9, 2015, https://doi.org/10.1007/s11033-015-3922-6
  10. Cloning, molecular characterization and functional analysis of 1-hydroxy-2-methyl-2-(E)-butenyl-4-diphosphate reductase (HDR) gene for diterpenoid tanshinone biosynthesis in Salvia miltiorrhiza Bge. f. alba vol.70, 2013, https://doi.org/10.1016/j.plaphy.2013.05.010
  11. In silico characterization and differential expression pattern analysis of conserved HMG CoA reductase domain isolated from Aconitum balfourii Stapf vol.6, pp.1, 2016, https://doi.org/10.1007/s13205-016-0405-y
  12. Isolation and characterization of a gene encoding 3-hydroxy-3-methylglutary coenzyme A reductase from an endophytic taxol-producing fungus BT2 vol.62, pp.2, 2012, https://doi.org/10.1007/s13213-011-0295-5
  13. Cloning and characterization of a novel 3-hydroxy-3-methylglutaryl coenzyme A reductase gene from Salvia miltiorrhiza involved in diterpenoid tanshinone accumulation vol.168, pp.2, 2011, https://doi.org/10.1016/j.jplph.2010.06.008
  14. A raison d’être for two distinct pathways in the early steps of plant isoprenoid biosynthesis? vol.51, pp.2, 2012, https://doi.org/10.1016/j.plipres.2011.12.001
  15. Molecular cloning, characterization and expression analysis of a new gene encoding 3-hydroxy-3-methylglutaryl coenzyme A reductase from Salvia miltiorrhiza vol.31, pp.3, 2009, https://doi.org/10.1007/s11738-008-0266-z
  16. Regulation of Banana Phytoene Synthase (MaPSY) Expression, Characterization and Their Modulation under Various Abiotic Stress Conditions vol.8, 2017, https://doi.org/10.3389/fpls.2017.00462
  17. Molecular cloning and functional expression analysis of a new gene encoding geranylgeranyl diphosphate synthase from hazel (Corylus avellana L. Gasaway) vol.37, pp.7, 2010, https://doi.org/10.1007/s11033-009-9935-2
  18. Highly active human pharmaceuticals in aquatic systems: A concept for their identification based on their mode of action vol.96, pp.3, 2010, https://doi.org/10.1016/j.aquatox.2009.11.021
  19. HMG-CoA reductase from Camphor Tulsi (Ocimum kilimandscharicum) regulated MVA dependent biosynthesis of diverse terpenoids in homologous and heterologous plant systems vol.8, pp.1, 2018, https://doi.org/10.1038/s41598-017-17153-z