Plant Molecular Biology

, Volume 57, Issue 6, pp 855–870

CaaX-prenyltransferases are essential for expression of genes involvedin the early stages of monoterpenoid biosynthetic pathwayin Catharanthus roseus cells

  • Vincent Courdavault
  • Martine Thiersault
  • Martine Courtois
  • Pascal Gantet
  • Audrey Oudin
  • Pierre Doireau
  • Benoit St-Pierre
  • Nathalie Giglioli-Guivarc’h
Article

DOI: 10.1007/s11103-005-3095-0

Cite this article as:
Courdavault, V., Thiersault, M., Courtois, M. et al. Plant Mol Biol (2005) 57: 855. doi:10.1007/s11103-005-3095-0

Abstract

CaaX-prenyltransferases (CaaX-PTases) catalyse the covalent attachment of isoprenyl groups to conserved cysteine residues located at the C-terminal CaaX motif of a protein substrate. This post-translational modification is required for the function and/or subcellular localization of some transcription factors and components of signal transduction and membrane trafficking machinery. CaaX-PTases, including protein farnesyltransferase (PFT) and type-I protein geranylgeranyltransferase (PGGT-I), are heterodimeric enzymes composed of a common α subunit and a specific β subunit. We have established RNA interference cell lines targeting the β subunits of PFT and PGGT-I, respectively, in the Catharanthus roseus C20D cell line, which synthesizes monoterpenoid indole alkaloids in response to auxin depletion from the culture medium. In both types of RNAi cell lines, expression of a subset of genes involved in the early stage of monoterpenoid biosynthetic pathway (ESMB genes), including the MEP pathway, is strongly decreased. The role of CaaX-PTases in ESMB gene regulation was confirmed by using the general prenyltransferase inhibitor s-perillyl alcohol (SP) and the specific PFT inhibitor Manumycin A on the wild type line. Furthermore, supplementation of SP inhibited cells with monoterpenoid intermediates downstream of the steps encoded by the ESMB genes restores monoterpenoid indole alkaloids biosynthesis. We conclude that protein targets for both PFT and PGGT-I are required for the expression of ESMB genes and monoterpenoid biosynthesis in C. roseus, this represents a non previously described role for protein prenyltransferase in plants.

Keywords

Catharanthus roseus MEP pathway monoterpene indole alkaloids protein farnesyltransferase protein type-I geranylgeranyltransferase RNA interference 

Abbreviations

ESMB genes

early stages of monoterpenoid biosynthetic pathway genes

MIA

monoterpenoid indole alkaloids

MM

maintenance medium

PFT

protein farnesyltransferase

PGGT-I

type-I protein geranylgeranyltransferase

PM

production medium

PPT

protein- prenyltransferase

Copyright information

© Springer 2005

Authors and Affiliations

  • Vincent Courdavault
    • 1
  • Martine Thiersault
    • 1
  • Martine Courtois
    • 2
  • Pascal Gantet
    • 1
  • Audrey Oudin
    • 2
  • Pierre Doireau
    • 1
  • Benoit St-Pierre
    • 1
  • Nathalie Giglioli-Guivarc’h
    • 1
  1. 1.«Biomolécules et Biotechnologies Végétales», Labaratoire de Physiologie Végétale, UFR Science et TechniquesUniversité François-Rabelais de ToursToursFrance
  2. 2.«Biomolécules et Biotechnologies Végétales», Laberatoire de Biologie Moléculaire et Biochimie Végétale, UFR Science PharmaceutiquesUniversité François-Rabelais de ToursToursFrance
  3. 3.«Equipe analyse fonctionelle du génome du Riz»Université Montpellier IIFrance

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