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

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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.

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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

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  1. Pascal Gantet

    Present address: «Equipe analyse fonctionelle du génome du Riz», Université Montpellier II, C.C. 002, 34095, Montpellier Cedex 5, France

Authors and Affiliations

  1. «Biomolécules et Biotechnologies Végétales», Labaratoire de Physiologie Végétale, UFR Science et Techniques, Université François-Rabelais de Tours, EA2106, 37200 , Parc de Grandmont, Tours, France

    Vincent Courdavault, Martine Thiersault, Pascal Gantet, Pierre Doireau, Benoit St-Pierre & Nathalie Giglioli-Guivarc’h

  2. «Biomolécules et Biotechnologies Végétales», Laberatoire de Biologie Moléculaire et Biochimie Végétale, UFR Science Pharmaceutiques, Université François-Rabelais de Tours, EA 2106, 37200 , 31 avenue Monge, Tours, France

    Martine Courtois & Audrey Oudin

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  1. Vincent Courdavault
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Correspondence to Nathalie Giglioli-Guivarc’h.

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Courdavault, V., Thiersault, M., Courtois, M. et al. CaaX-prenyltransferases are essential for expression of genes involvedin the early stages of monoterpenoid biosynthetic pathwayin Catharanthus roseus cells. Plant Mol Biol 57, 855–870 (2005). https://doi.org/10.1007/s11103-005-3095-0

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