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Molecular cloning and functional characterization of Catharanthus roseus hydroxymethylbutenyl 4-diphosphate synthase gene promoter from the methyl erythritol phosphate pathway.
Mol Biol Rep. 2012 May; 39(5):5433-47.MB

Abstract

The Madagascar periwinkle produces monoterpenoid indole alkaloids (MIA) of high interest due to their therapeutical values. The terpenoid moiety of MIA is derived from the methyl erythritol phosphate (MEP) and seco-iridoid pathways. These pathways are regarded as the limiting branch for MIA biosynthesis in C. roseus cell and tissue cultures. In previous studies, we demonstrated a coordinated regulation at the transcriptional and spatial levels of genes from both pathways. We report here on the isolation of the 5'-flanking region (1,049 bp) of the hydroxymethylbutenyl 4-diphosphate synthase (HDS) gene from the MEP pathway. To investigate promoter transcriptional activities, the HDS promoter was fused to GUS reporter gene. Agrobacterium-mediated transformation of young tobacco leaves revealed that the cloned HDS promoter displays a tissue-specific GUS staining restricted to the vascular region of the leaves and limited to a part of the vein that encompasses the phloem in agreement with the previous localization of HDS transcripts in C. roseus aerial organs. Further functional characterizations in stably or transiently transformed C. roseus cells allowed us to identify the region that can be consider as the minimal promoter and to demonstrate the induction of HDS promoter by several hormonal signals (auxin, cytokinin, methyljasmonate and ethylene) leading to MIA production. These results, and the bioinformatic analysis of the HDS 5'-region, suggest that the HDS promoter harbours a number of cis-elements binding specific transcription factors that would regulate the flux of terpenoid precursors involved in MIA biosynthesis.

Authors+Show Affiliations

Université François Rabelais de Tours, EA 2106, Biomolécules et Biotechnologies Végétales, 31 avenue Monge, 37200, Tours, France.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

22160472

Citation

Ginis, Olivia, et al. "Molecular Cloning and Functional Characterization of Catharanthus Roseus Hydroxymethylbutenyl 4-diphosphate Synthase Gene Promoter From the Methyl Erythritol Phosphate Pathway." Molecular Biology Reports, vol. 39, no. 5, 2012, pp. 5433-47.
Ginis O, Courdavault V, Melin C, et al. Molecular cloning and functional characterization of Catharanthus roseus hydroxymethylbutenyl 4-diphosphate synthase gene promoter from the methyl erythritol phosphate pathway. Mol Biol Rep. 2012;39(5):5433-47.
Ginis, O., Courdavault, V., Melin, C., Lanoue, A., Giglioli-Guivarc'h, N., St-Pierre, B., Courtois, M., & Oudin, A. (2012). Molecular cloning and functional characterization of Catharanthus roseus hydroxymethylbutenyl 4-diphosphate synthase gene promoter from the methyl erythritol phosphate pathway. Molecular Biology Reports, 39(5), 5433-47. https://doi.org/10.1007/s11033-011-1343-8
Ginis O, et al. Molecular Cloning and Functional Characterization of Catharanthus Roseus Hydroxymethylbutenyl 4-diphosphate Synthase Gene Promoter From the Methyl Erythritol Phosphate Pathway. Mol Biol Rep. 2012;39(5):5433-47. PubMed PMID: 22160472.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Molecular cloning and functional characterization of Catharanthus roseus hydroxymethylbutenyl 4-diphosphate synthase gene promoter from the methyl erythritol phosphate pathway. AU - Ginis,Olivia, AU - Courdavault,Vincent, AU - Melin,Céline, AU - Lanoue,Arnaud, AU - Giglioli-Guivarc'h,Nathalie, AU - St-Pierre,Benoit, AU - Courtois,Martine, AU - Oudin,Audrey, Y1 - 2011/12/13/ PY - 2011/07/06/received PY - 2011/12/03/accepted PY - 2011/12/14/entrez PY - 2011/12/14/pubmed PY - 2012/7/13/medline SP - 5433 EP - 47 JF - Molecular biology reports JO - Mol Biol Rep VL - 39 IS - 5 N2 - The Madagascar periwinkle produces monoterpenoid indole alkaloids (MIA) of high interest due to their therapeutical values. The terpenoid moiety of MIA is derived from the methyl erythritol phosphate (MEP) and seco-iridoid pathways. These pathways are regarded as the limiting branch for MIA biosynthesis in C. roseus cell and tissue cultures. In previous studies, we demonstrated a coordinated regulation at the transcriptional and spatial levels of genes from both pathways. We report here on the isolation of the 5'-flanking region (1,049 bp) of the hydroxymethylbutenyl 4-diphosphate synthase (HDS) gene from the MEP pathway. To investigate promoter transcriptional activities, the HDS promoter was fused to GUS reporter gene. Agrobacterium-mediated transformation of young tobacco leaves revealed that the cloned HDS promoter displays a tissue-specific GUS staining restricted to the vascular region of the leaves and limited to a part of the vein that encompasses the phloem in agreement with the previous localization of HDS transcripts in C. roseus aerial organs. Further functional characterizations in stably or transiently transformed C. roseus cells allowed us to identify the region that can be consider as the minimal promoter and to demonstrate the induction of HDS promoter by several hormonal signals (auxin, cytokinin, methyljasmonate and ethylene) leading to MIA production. These results, and the bioinformatic analysis of the HDS 5'-region, suggest that the HDS promoter harbours a number of cis-elements binding specific transcription factors that would regulate the flux of terpenoid precursors involved in MIA biosynthesis. SN - 1573-4978 UR - https://www.unboundmedicine.com/medline/citation/22160472/Molecular_cloning_and_functional_characterization_of_Catharanthus_roseus_hydroxymethylbutenyl_4_diphosphate_synthase_gene_promoter_from_the_methyl_erythritol_phosphate_pathway_ L2 - https://doi.org/10.1007/s11033-011-1343-8 DB - PRIME DP - Unbound Medicine ER -