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Differential expression of the TwHMGS gene and its effect on triptolide biosynthesis in Tripterygium wilfordii
Alternative TitleDifferential expression of the TwHMGS gene and its effect on triptolide biosynthesis in Tripterygium wilfordii
Tong YuRu1; Zhang YiFeng3; Zhao YuJun2; Hu TianYuan3; Wang JiaDian3; Huang LuQi1; Gao Wei3
2019
Source PublicationCHINESE JOURNAL OF NATURAL MEDICINES
ISSN2095-6975
Volume17Issue:8Pages:575-584
Abstract3-Hydroxy-3-methylglutaryl-CoA synthase (HMGS) is the first committed enzyme in the MVA pathway and involved in the biosynthesis of terpenes in Tripterygium wilfordii. The full- length cDNA and a 515 bp RNAi target fragment of TwHMGS were ligated into the pH7WG2D and pK7GWIWG2D vectors to respectively overexpress and silence, TwHMGS was overexpressed and silenced in T. wilfordii suspension cells using biolistic-gun mediated transformation, which resulted in 2-fold increase and a drop to 70% in the expression level compared to cells with empty vector controls. During TwHMGS overexpression, the expression of TwHMGR, TwDXR and TwTPS7v2 was significantly upregulated to the control. In the RNAi group, the expression of TwHMGR, TwDXS, TwDXR and TwMCT visibly displayed downregulation to the control. The cells with TwHMGS overexpressed produced twice higher than the control value. These results proved that differential expression of TwHMGS determined the production of triptolide in T. wilfordii and laterally caused different trends of relative gene expression in the terpene biosynthetic pathway. Finally, the substrate acetyl-CoA was docked into the active site of TwHMGS, suggesting the key residues including His247, Lys256 and Arg296 undergo electrostatic or H-bond interactions with acetyl-CoA.
Other Abstract3-Hydroxy-3-methylglutaryl-CoA synthase (HMGS) is the first committed enzyme in the MVA pathway and involved in the biosynthesis of terpenes in Tripterygium wilfordii. The full- length cDNA and a 515 bp RNAi target fragment of TwHMGS were ligated into the pH7WG2D and pK7GWIWG2D vectors to respectively overexpress and silence, TwHMGS was overexpressed and silenced in T. wilfordii suspension cells using biolistic-gun mediated transformation, which resulted in 2-fold increase and a drop to 70% in the expression level compared to cells with empty vector controls. During TwHMGS overexpression, the expression of TwHMGR, TwDXR and TwTPS7v2 was significantly upregulated to the control. In the RNAi group, the expression of TwHMGR, TwDXS, TwDXR and TwMCT visibly displayed downregulation to the control. The cells with TwHMGS overexpressed produced twice higher than the control value. These results proved that differential expression of TwHMGS determined the production of triptolide in T. wilfordii and laterally caused different trends of relative gene expression in the terpene biosynthetic pathway. Finally, the substrate acetyl-CoA was docked into the active site of TwHMGS, suggesting the key residues including His247, Lys256 and Arg296 undergo electrostatic or H-bond interactions with acetyl-CoA.
KeywordENCODING 3-HYDROXY-3-METHYLGLUTARYL-COA SYNTHASE HMG-COA SYNTHASE METHYLERYTHRITOL PHOSPHATE ISOPRENOID BIOSYNTHESIS MOLECULAR-CLONING CROSS-TALK WILD-TYPE PATHWAYS OVEREXPRESSION INTERMEDIATE Overexpression RNAi HMGS Triptolide Acetyl-CoA
Indexed ByCSCD
Language英语
Funding Project[National Natural Science Foundation of China] ; [Beijing Natural Science Foundation Program] ; [Scientific Research Key Program of Beijing Municipal Commission of Education] ; [Support Project of High-level Teachers in Beijing Municipal Universities in the Period of 13th Five-year Plan] ; [National Program for Special Support of Eminent Professionals] ; [Key project at central government level: The ability establishment of sustainable use for valuable Chinese medicine resources]
CSCD IDCSCD:6543711
Citation statistics
Cited Times:1[CSCD]   [CSCD Record]
Document Type期刊论文
Identifierhttp://ir.imr.ac.cn/handle/321006/145242
Collection中国科学院金属研究所
Affiliation1.中国科学院金属研究所
2.中国科学院
3.Capital Med University, Sch Pharmaceut Sci, Beijing 100069, Peoples R China
4.Capital Med University, Sch Tradit Chinese Med, Beijing 100069, Peoples R China Sch Tradit Chinese Med
5.Capital Med University, Adv Innovat Ctr Human Brain Protect, Beijing 100069, Peoples R China
Recommended Citation
GB/T 7714
Tong YuRu,Zhang YiFeng,Zhao YuJun,et al. Differential expression of the TwHMGS gene and its effect on triptolide biosynthesis in Tripterygium wilfordii[J]. CHINESE JOURNAL OF NATURAL MEDICINES,2019,17(8):575-584.
APA Tong YuRu.,Zhang YiFeng.,Zhao YuJun.,Hu TianYuan.,Wang JiaDian.,...&Gao Wei.(2019).Differential expression of the TwHMGS gene and its effect on triptolide biosynthesis in Tripterygium wilfordii.CHINESE JOURNAL OF NATURAL MEDICINES,17(8),575-584.
MLA Tong YuRu,et al."Differential expression of the TwHMGS gene and its effect on triptolide biosynthesis in Tripterygium wilfordii".CHINESE JOURNAL OF NATURAL MEDICINES 17.8(2019):575-584.
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