miR5298b regulated taxol biosynthesis by acting on TcNPR3, resulting in an alleviation of the strong inhibition of the TcNPR3-TcTGA6 complex in Taxus chinensis

Chen Ying; Zhang Meng; Zhang Wenli; Wang Yamin; Zhang Hua; Yang Liu; Yu Longjiang; Fu Chunhua

doi:10.1016/j.ijbiomac.2023.125909

miR5298b regulated taxol biosynthesis by acting on TcNPR3, resulting in an alleviation of the strong inhibition of the TcNPR3-TcTGA6 complex in Taxus chinensis

Int J Biol Macromol. 2023 Sep 1:248:125909. doi: 10.1016/j.ijbiomac.2023.125909. Epub 2023 Jul 22.

Authors

Chen Ying¹, Zhang Meng², Zhang Wenli¹, Wang Yamin¹, Zhang Hua¹, Yang Liu³, Yu Longjiang¹, Fu Chunhua⁴

Affiliations

¹ Institute of Resource Biology and Biotechnology, Department of Biotechnology, College of Life Science and Technology, Huazhong University of Science and Technology, No.1037 Luoyu Road, Wuhan 430074, PR China; Key Laboratory of Molecular Biophysics Ministry of Education, College of Life Science and Technology, Huazhong University of Science and Technology, No.1037 Luoyu Road, Wuhan 430074, PR China; Hubei Food and medicine Resources Engineering Research Center, College of Life Science and Technology, Huazhong University of Science and Technology, No.1037 Luoyu Road, Wuhan 430074, PR China.
² Institute of Resource Biology and Biotechnology, Department of Biotechnology, College of Life Science and Technology, Huazhong University of Science and Technology, No.1037 Luoyu Road, Wuhan 430074, PR China; Hunan Key Laboratory of Plant Functional Genomics and Developmental Regulation, College of Biology, Hunan University, Changsha, 410082, PR China.
³ Hunan Key Laboratory of Plant Functional Genomics and Developmental Regulation, College of Biology, Hunan University, Changsha, 410082, PR China.
⁴ Institute of Resource Biology and Biotechnology, Department of Biotechnology, College of Life Science and Technology, Huazhong University of Science and Technology, No.1037 Luoyu Road, Wuhan 430074, PR China; Key Laboratory of Molecular Biophysics Ministry of Education, College of Life Science and Technology, Huazhong University of Science and Technology, No.1037 Luoyu Road, Wuhan 430074, PR China; Hubei Food and medicine Resources Engineering Research Center, College of Life Science and Technology, Huazhong University of Science and Technology, No.1037 Luoyu Road, Wuhan 430074, PR China. Electronic address: fuchunhua@mail.hust.edu.cn.

PMID: 37482165
DOI: 10.1016/j.ijbiomac.2023.125909

Abstract

Taxol, a valuable but rare secondary metabolite of the genus Taxus, is an effective anticancer drug. Understanding the regulation of taxol biosynthesis may provide a means to increase taxol content. The microRNA miR5298b was found to promote the accumulation of taxol and upregulate several taxol biosynthesis genes, including DBAT, TASY, and T5H, as demonstrated by experiments using the overexpression and mimicry of transient leaves. Moreover, miR5298b cleaves the mRNA sequence of TcNPR3, a homolog of the salicylic acid receptor AtNPR3/4. Overexpression and knockdown by RNA interference of TcNPR3 confirmed that it repressed taxol biosynthesis. These results indicate that miR5298b enhances taxol biosynthesis via the cleavage of TcNPR3. Yeast two-hybrid bimolecular fluorescence complementation and pull-down assays revealed that TcTGA6, a TGA transcription factor, physically interacted with TcNPR3. Functional experiments showed that TcTGA6 negatively regulates taxol biosynthesis by directly combining with the TGACG motif in the promoters of TASY, T5H, and T10H. TcNPR3 enhances TcTGA6 inhibition Luciferase assays showed that miR5298b alleviated the repression of the TcNPR3-TcTGA6 complex. In summary, miR5298b can cleave TcNPR3, thereby alleviating the inhibition of the TcNPR3-TcTGA6 complex to upregulate taxol biosynthesis genes.

Keywords: TGA; Taxol biosynthesis; Taxus chinensis; TcNPR3; miR5298b.

MeSH terms

MicroRNAs* / genetics
MicroRNAs* / metabolism
Paclitaxel / metabolism
Promoter Regions, Genetic / genetics
Taxus* / genetics
Taxus* / metabolism
Transcription Factors / genetics

Substances

Transcription Factors
Paclitaxel
MicroRNAs