A pair of tabersonine 16-hydroxylases initiates the synthesis of vindoline in an organ-dependent manner in Catharanthus roseus
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A pair of tabersonine 16-hydroxylases initiates the synthesis of vindoline in an organ-dependent manner in Catharanthus roseus. / Besseau, Sébastien; Kellner, Franziska; Lanoue, Arnaud; Thamm, Antje M K; Salim, Vonny; Schneider, Bernd; Geu Flores, Fernando; Höfer, René; Guirimand, Grégory; Guihur, Anthony; Oudin, Audrey; Glevarec, Gaëlle; Foureau, Emilien; Papon, Nicolas; Clastre, Marc; Giglioli-Guivarc'h, Nathalie; St-Pierre, Benoit; Werck-Reichhart, Danièle; Burlat, Vincent; De Luca, Vincenzo; O'Connor, Sarah E; Courdavault, Vincent.
In: Plant Physiology, Vol. 163, No. 4, 2013, p. 1792-803.Research output: Contribution to journal › Journal article › Research › peer-review
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TY - JOUR
T1 - A pair of tabersonine 16-hydroxylases initiates the synthesis of vindoline in an organ-dependent manner in Catharanthus roseus
AU - Besseau, Sébastien
AU - Kellner, Franziska
AU - Lanoue, Arnaud
AU - Thamm, Antje M K
AU - Salim, Vonny
AU - Schneider, Bernd
AU - Geu Flores, Fernando
AU - Höfer, René
AU - Guirimand, Grégory
AU - Guihur, Anthony
AU - Oudin, Audrey
AU - Glevarec, Gaëlle
AU - Foureau, Emilien
AU - Papon, Nicolas
AU - Clastre, Marc
AU - Giglioli-Guivarc'h, Nathalie
AU - St-Pierre, Benoit
AU - Werck-Reichhart, Danièle
AU - Burlat, Vincent
AU - De Luca, Vincenzo
AU - O'Connor, Sarah E
AU - Courdavault, Vincent
PY - 2013
Y1 - 2013
N2 - Hydroxylation of tabersonine at the C-16 position, catalyzed by tabersonine 16-hydroxylase (T16H), initiates the synthesis of vindoline that constitutes the main alkaloid accumulated in leaves of Catharanthus roseus. Over the last decade, this reaction has been associated with CYP71D12 cloned from undifferentiated C. roseus cells. In this study, we isolated a second cytochrome P450 (CYP71D351) displaying T16H activity. Biochemical characterization demonstrated that CYP71D12 and CYP71D351 both exhibit high affinity for tabersonine and narrow substrate specificity, making of T16H, to our knowledge, the first alkaloid biosynthetic enzyme displaying two isoforms encoded by distinct genes characterized to date in C. roseus. However, both genes dramatically diverge in transcript distribution in planta. While CYP71D12 (T16H1) expression is restricted to flowers and undifferentiated cells, the CYP71D351 (T16H2) expression profile is similar to the other vindoline biosynthetic genes reaching a maximum in young leaves. Moreover, transcript localization by carborundum abrasion and RNA in situ hybridization demonstrated that CYP71D351 messenger RNAs are specifically located to leaf epidermis, which also hosts the next step of vindoline biosynthesis. Comparison of high- and low-vindoline-accumulating C. roseus cultivars also highlights the direct correlation between CYP71D351 transcript and vindoline levels. In addition, CYP71D351 down-regulation mediated by virus-induced gene silencing reduces vindoline accumulation in leaves and redirects the biosynthetic flux toward the production of unmodified alkaloids at the C-16 position. All these data demonstrate that tabersonine 16-hydroxylation is orchestrated in an organ-dependent manner by two genes including CYP71D351, which encodes the specific T16H isoform acting in the foliar vindoline biosynthesis.
AB - Hydroxylation of tabersonine at the C-16 position, catalyzed by tabersonine 16-hydroxylase (T16H), initiates the synthesis of vindoline that constitutes the main alkaloid accumulated in leaves of Catharanthus roseus. Over the last decade, this reaction has been associated with CYP71D12 cloned from undifferentiated C. roseus cells. In this study, we isolated a second cytochrome P450 (CYP71D351) displaying T16H activity. Biochemical characterization demonstrated that CYP71D12 and CYP71D351 both exhibit high affinity for tabersonine and narrow substrate specificity, making of T16H, to our knowledge, the first alkaloid biosynthetic enzyme displaying two isoforms encoded by distinct genes characterized to date in C. roseus. However, both genes dramatically diverge in transcript distribution in planta. While CYP71D12 (T16H1) expression is restricted to flowers and undifferentiated cells, the CYP71D351 (T16H2) expression profile is similar to the other vindoline biosynthetic genes reaching a maximum in young leaves. Moreover, transcript localization by carborundum abrasion and RNA in situ hybridization demonstrated that CYP71D351 messenger RNAs are specifically located to leaf epidermis, which also hosts the next step of vindoline biosynthesis. Comparison of high- and low-vindoline-accumulating C. roseus cultivars also highlights the direct correlation between CYP71D351 transcript and vindoline levels. In addition, CYP71D351 down-regulation mediated by virus-induced gene silencing reduces vindoline accumulation in leaves and redirects the biosynthetic flux toward the production of unmodified alkaloids at the C-16 position. All these data demonstrate that tabersonine 16-hydroxylation is orchestrated in an organ-dependent manner by two genes including CYP71D351, which encodes the specific T16H isoform acting in the foliar vindoline biosynthesis.
KW - Biocatalysis
KW - Biosynthetic Pathways
KW - Catharanthus
KW - Cytochrome P-450 Enzyme System
KW - DNA, Complementary
KW - Endoplasmic Reticulum
KW - Gene Expression Regulation, Enzymologic
KW - Gene Expression Regulation, Plant
KW - Gene Silencing
KW - Genes, Plant
KW - Hydroxylation
KW - Indole Alkaloids
KW - Kinetics
KW - Metabolome
KW - Molecular Sequence Data
KW - Organ Specificity
KW - Plant Epidermis
KW - Plant Proteins
KW - Quinolines
KW - RNA, Messenger
KW - Substrate Specificity
KW - Vinblastine
U2 - 10.1104/pp.113.222828
DO - 10.1104/pp.113.222828
M3 - Journal article
C2 - 24108213
VL - 163
SP - 1792
EP - 1803
JO - Plant Physiology
JF - Plant Physiology
SN - 0032-0889
IS - 4
ER -
ID: 130370647