Fig. 3: Characterization of FoTO1.
From: Discovery of FoTO1 and Taxol genes enables biosynthesis of baccatin III
a, Early Taxol biosynthetic pathway and the T5αH product divergence. Blue shading highlights the biosynthetic pathway towards Taxol. Diamond indicates the structure is supported by NMR. b, Rank and score of genes in Taxol module 1. c, Bar graph showing the FoTO1-induced fold change in end-products’ peak area of subpathways when transiently expressed in N. benthamiana leaves. Fold change is calculated by quantifying the GC–MS total ion chromatogram (TIC) peak area of compounds 2–4 and normalizing to the −FoTO1 condition. cytoTDS, cytosolic TDS. Data are mean ± s.d., n = 3 biological, independent leaf samples. Statistical analyses were performed using a two-sided, unpaired Welch’s t-test. d, GC–MS TIC of N. benthamiana leaves transiently expressing the indicated genes. e, Bar graph of total oxidized taxanes for the +T5αH and +T5αΗ+FoTO1 conditions. Data are mean ± s.d. n = 6 biological, independent leaf samples. One asterisk indicates previously characterized; two asterisks indicate characterized in this study. f, Phylogenetic tree of FoTO1 homologues identified by HMMER. The tree was produced with FastTree, rooted with red algae homologues. g, Structural model of FoTO1, generated by AlphaFold3 and aligned with the Arabidopsis thaliana orthologue with FoldSeek. h, Bar graphs showing the integrated peak area of 2 and 2’a when N- or C-terminally truncated FoTO1 is transiently expressed in N. benthamiana leaves together with TDS and T5αH. Data are mean ± s.d., n = 3 biological replicates. i, Quantification of binding between purified T5αΗ and FoTO1 or FoTO1(ΔC) using microscale thermophoresis. Data are mean ± s.d., n = 3 replicates. j, Quantification of binding between purified TDS2 and FoTO1 or FoTO1(ΔC), as in i (n = 3 replicates). N-terminal transmembrane domains of T5αΗ and TDS2 are removed for purification purposes. Data are mean ± s.d., n = 3. k, Immunoblot of the co-IP of T5αH–HA (prey) by V5–FoTO1 (bait) in N. benthamiana leaves expressing both proteins (Supplementary Fig. 7).
