Abstract
Paclitaxel is an important anticancer drug. The phytohormone jasmonic acid can significantly induce the biosynthesis of paclitaxel in Taxus, but the molecular mechanism has not yet been resolved. To establish the jasmonic acid signalling pathway of Taxus media, based on the gene of the jasmonic acid signalling pathway of Arabidopsis thaliana, sequence analysis was performed to isolate the jasmonic acid signal from the transcriptome, a transcriptional cluster of pathway gene homologs and the full length of 22 genes were obtained by RACE PCR at 5′ and 3′: two EI ubiquitin ligase genes, COI1-1 and COI1-2;7 MYC bHLH type transcription factor (MYC2, MYC3, MYC4, JAM1, JAM2, EGL3, TT8); 12 JAZ genes containing the ZIM domain; and MED25, one of the components of the transcriptional complex. The protein interaction between each were confirmed by yeast two hybridization and bimolecular fluorescence complementation based on similar genes interaction in Arabidopsis. A similar jasmonate signaling pathway was illustrated in T. media. All known paclitaxel biosynthesis genes promoters were isolated by genome walker PCR. To investigate the jasmonate signaling effect on these genes’ expression, the transcription activity of MYC2, MYC3 and MYC4 on these promoters were examined. There are 12, 10 and 11 paclitaxel biosynthesis genes promoters that could be activated by MYC2, MYC3 and MYC4.
Highlights
Paclitaxel is an important anticancer drug, which is mainly produced by chemical semi-synthesis by using paclitaxel, which is biosynthesized in Pacific yew
We found that some of the genes involved in the paclitaxel synthesis pathway were highly expressed by jasmonic acid at 0.5 h, similar to the transcription factors of the jasmonic acid signalling pathway and the expression of self-synthesis-related genes, but faster
For comprehensive detection of JAZ that may be contained in the transcriptome, BLAST analysis was performed using the full length of the Arabidopsis AtJAZ1 gene and the eight JAZs with reference to the Tm_ZK transcriptome, and the threshold of the E value was set to 0.01
Summary
Paclitaxel is an important anticancer drug, which is mainly produced by chemical semi-synthesis by using paclitaxel, which is biosynthesized in Pacific yew. The phytohormone jasmonic acid can significantly induce the biosynthesis of paclitaxel in Taxus, but the molecular mechanism has not yet been resolved [2,3,4]. The synthetic pathway can be divided into the following four steps: taxane ring synthesis; side hydroxylation, acetylation and phenylacetylation modification; epoxidation modification at C5 position and oxidation at C9; and side chain at C13 position addition [5]. The precursor of taxadiene ring biosynthesis is derived from isopentenyl pyrophosphate (IPP) and dimethyl propylene pyrophosphate (DMAPP), the synthesis, mainly through the plastids of Taxus cells [5,6,7]. After ring formation, the biosynthesis process of paclitaxel includes various modifications to the sites on the ring, including hydroxylation, acetylation and benzoylation modification, epoxidation at the C5 position, and oxidative modification at the C9 position [7]
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