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Identification of two MYB transcription factors that increase paclitaxel biosynthesis in cambial meristematic cells of Taxus baccata

Authors: Ochoa-Villarreal, Marisol;

Identification of two MYB transcription factors that increase paclitaxel biosynthesis in cambial meristematic cells of Taxus baccata

Abstract

Paclitaxel is an anticancer natural product with several biomedical applications produced by Taxus species, with a demand exceeding its supply. We have developed cambial meristematic cells (CMCs) from Taxus cuspidata as high yield source of paclitaxel. The biosynthesis of paclitaxel is predominantly under transcriptional control. Thus, the identification of transcriptional regulators of paclitaxel biosynthesis and their subsequent manipulation may enable further yield enhancement in Taxus CMCs. Previously, Roche 454 sequencing was employed to establish the transcriptome of T. cuspidata CMCs treated with the plant immune activator methyl jasmonate (MeJA). The bioinformatic analysis identified 19 jasmonate related transcription factors (TFs), based on their differential expression. Results of the Arabidopsis thaliana transient assay screen identified two MYB TFs that constitute positive regulators for paclitaxel genes, named MYB3 and MYB4. In this thesis, MYB3 and MYB4 showed in vitro binding to the cis-elements in ten promoters of paclitaxel genes using the electrophoretic mobility shift assay (EMSA). Then, a Taxus CMC protoplasts transient assay demonstrated that the expression of MYB3 and MYB4 trans-activated all tested genes. Further, MYB4 was found to activate the 3-hydroxy-3-methylglutaryl-CoA (HMG-CoA) gene, key in the mevalonate pathway and precursor of paclitaxel biosynthesis. MYB3 and MYB4 were capable of auto-regulating their own transcription, constituting an important control point for paclitaxel biosynthesis. A possible mechanism for the early activation of MYB3 and MYB4 after MeJA elicitation is proposed. Finally, preliminary results on the expression of MYB3 and MYB4 in unelicited T. baccata CMC protoplasts indicate that their transient expression was sufficient to increase accumulation of paclitaxel and the precursor, 10-deacetyl baccatin III, highlighting their utility for paclitaxel production.

Country
United Kingdom
Related Organizations
Keywords

plant natural products, Paclitaxel, plant cell culture, transcription factors, cambial meristematic cells, anticancer

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selected citations
These citations are derived from selected sources.
This is an alternative to the "Influence" indicator, which also reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Citations provided by BIP!
popularity
This indicator reflects the "current" impact/attention (the "hype") of an article in the research community at large, based on the underlying citation network.
BIP!Popularity provided by BIP!
influence
This indicator reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Influence provided by BIP!
impulse
This indicator reflects the initial momentum of an article directly after its publication, based on the underlying citation network.
BIP!Impulse provided by BIP!
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