title: Two jasmonate-responsive factors, TcERF12 and TcERF15, respectively act as repressor and activator of tasy gene of taxol biosynthesis in Taxus chinensis.
abstract: Methyl jasmonate (MeJA) is one of the most effective inducers of taxol biosynthetic genes, particularly the tasy gene. However, the mechanism underlying the regulation of tasy by MeJA is still unknown. In this study, a 550-bp 5'-flanking sequence was obtained and confirmed as the promoter of the tasy gene. Deletion analysis revealed that the fragment containing a GCC-box from -150 to -131 was the crucial jasmonate (JA)-responsive element, designated as JRE. Using JRE as bait, two binding proteins, namely TcERF12 and TcERF15, were discovered. Sequence alignment and phylogenetic analysis showed that TcERF12 was related to the repressor AtERF3, while TcERF15 was more related to the activator ORA59; these are typical GCC-box-binding ethylene-responsive factors. Both could significantly respond to MeJA for 10 and 4.5 times, respectively, in 0.5 h. When the two TcERFs were overexpressed in Taxus cells, tasy gene expression decreased by 2.1 times in TcERF12-overexpressing cells, but increased by 2.5 times in TcERF15-overexpressing cells. Results indicated that TcERF12 and TcERF15 were negative and positive regulators, respectively, in the JA signal transduction to the tasy gene by binding the GCC-box in the JRE of the tasy promoter. Our results promote further research on regulatory mechanisms of taxol biosynthesis.
authors.affiliation: 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, People's Republic of 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, People's Republic of China.
authors.forename: Meng
authors.initials: M
authors.lastname: Zhang
authors.affiliation: 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, People's Republic of 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, People's Republic of China.
authors.forename: Shutao
authors.initials: S
authors.lastname: Li
authors.affiliation: 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, People's Republic of 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, People's Republic of China.
authors.forename: Lin
authors.initials: L
authors.lastname: Nie
authors.affiliation: 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, People's Republic of 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, People's Republic of China.
authors.forename: Qingpu
authors.initials: Q
authors.lastname: Chen
authors.affiliation: 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, People's Republic of 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, People's Republic of China.
authors.forename: Xiangping
authors.initials: X
authors.lastname: Xu
authors.affiliation: 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, People's Republic of China. yulongjiang@mail.hust.edu.cn.|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, People's Republic of China. yulongjiang@mail.hust.edu.cn.
authors.forename: Longjiang
authors.initials: L
authors.lastname: Yu
authors.affiliation: 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, People's Republic of China. fuch2003@126.com.|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, People's Republic of China. fuch2003@126.com.
authors.forename: Chunhua
authors.initials: C
authors.lastname: Fu
mesh_terms: D024506:5' Flanking Region; D000595:Amino Acid Sequence; D001483:Base Sequence; D003517:Cyclopentanes / Q000378:metabolism*; D018744:DNA, Plant / Q000235:genetics; D018506:Gene Expression Regulation, Plant; D017343:Genes, Plant; D008969:Molecular Sequence Data; D054883:Oxylipins / Q000378:metabolism*; D017239:Paclitaxel / Q000096:biosynthesis*; D010802:Phylogeny; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism; D030821:Plants, Genetically Modified; D011401:Promoter Regions, Genetic; D017386:Sequence Homology, Amino Acid; D015398:Signal Transduction; D020946:Taxus / Q000235:genetics* / Q000378:metabolism*; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D020798:Two-Hybrid System Techniques
chemical_list: D003517:Cyclopentanes; D018744:DNA, Plant; D054883:Oxylipins; D010940:Plant Proteins; D014157:Transcription Factors; C011006:jasmonic acid; D017239:Paclitaxel
keywords: ERFs; GCC-box; MeJA; Tasy gene promoter; Taxus chinensis