Plant Molecular Biology

, Volume 57, Issue 6, pp 855–870 | Cite as

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


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.


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


ESMB genes

early stages of monoterpenoid biosynthetic pathway genes


monoterpenoid indole alkaloids


maintenance medium


protein farnesyltransferase


type-I protein geranylgeranyltransferase


production medium


protein- prenyltransferase


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  1. Arvy, M.P., Imbault, N., Naudascher, F., Thiersault, M., Doireau, P 19942,4-D and alkaloid accumulation in periwinkle cell suspensionsBiochimie76410416CrossRefPubMedGoogle Scholar
  2. Bach, T.J 1995Some new aspects of isoprenoid biosynthesis in plantsLipids30191202PubMedGoogle Scholar
  3. Burlat, V., Oudin, A., Courtois, M., Rideau, M., St-Pierre, B 2004Co-expression of three MEP pathway genes and geraniol 10-hydroxylase in internal phloem parenchyma of Catharanthus roseus implicates multicellular translocation of intermediates during the biosynthesis of monoterpene indole alkaloids and isoprenoid-derived primary metabolitesPlant J.38131141CrossRefPubMedGoogle Scholar
  4. Caldelari, D., Sternberg, H., Rodriguez-Concepcion, M., Gruissem, W., Yalovsky, S 2001Efficient prenylation by a plant geranylgeranyltransferase-I requires a functional CaaL box motif and a proximal polybasic domainPlant Physiol.12614161429CrossRefPubMedGoogle Scholar
  5. Casey, P.J 1995Protein lipidation in cell signallingScience268221225PubMedGoogle Scholar
  6. Casey, P.J., Solski, P.A., Der, C.J., Buss, J.E 1989p21ras is modified by a farnesyl isoprenoidProc. Natl. Acad. Sci. USA8683238327PubMedGoogle Scholar
  7. Chahed, K., Oudin, A., Guivarc’h, N., Hamdi, S., Chénieux, J.-C., Rideau, M., Clastre, M 20001-Deoxy-D-xylulose 5-phosphate synthase from periwinkle: cDNA identification and induced gene expression in terpenoid indole alkaloid-producing cellsPlant Physiol. Biochem.38559566CrossRefGoogle Scholar
  8. Chappell, J. 2002The genetics and molecular genetics of terpene and sterol origamiCurr. Opin. Plant Biol.5151157CrossRefPubMedGoogle Scholar
  9. Contin, A., Heijden, R., Lefeber, A.W.M., Verpoort, R 1998The iridoid glucoside secologanin is derived from the novel triose phosphate/pyruvate pathway in a Catharanthus roseus cell cultureFEBS Lett.437413416Google Scholar
  10. Courdavault, V., Burlat, V., St-Pierre, B. 2005Giglioli-Guivarc’h,N. Characterisation of CaaX-prenyltransferases in Catharanthus roseus: relationships with the expression of genes involved in the early stages of monoterpenoid biosynthetic pathwayPlant Sci.16810971107Google Scholar
  11. Crowell, P.L., Ren, Z., Lin, S., Vedejs, E., Gould, M.N 1994Structure-activity relationships among monoterpene inhibitors of protein isoprenylation and cell proliferationBiochem. Pharmacol.4714051415PubMedGoogle Scholar
  12. Cutler, S., Ghassemian, M., Bonetta, D., Cooney, S., McCourt, P 1996A protein farnesyltransferase involved in abscisic acid signal transduction in ArabidopsisScience27312391241PubMedGoogle Scholar
  13. Dellaporta, S.L., Wood, J., Hicks, J.B 1983A plant DNA minipreparation, Version IIPlant Mol. Biol. Rep.11921Google Scholar
  14. Farnsworth, C.C., Gelb, M.H., Glomset, J.A 1990Identification of geranylgeranyl-modified proteins in HeLa cellsScience247320322PubMedGoogle Scholar
  15. Gamborg, O.L., Miller, R.R.A., Ojima, K 1968Nutriment requirements of suspension cultures of soybean root cellsExp. Cells Res.50151158Google Scholar
  16. Gelb, M.H., Tamanoi, F., Yokoyama, K., Ghomashchi, F., Esson, K., Gould, M.N 1995The inhibition of protein prenyltransferases by oxygenated metabolites of limonene and perillyl alcoholCancer Lett.91169175PubMedGoogle Scholar
  17. Gibbs, J.B., Pompliano, D.L., Mosser, S.D., Rands, E., Lingham, R.B., Singh, S.B., Scolnick, E.M., Kohl, N.E., Oliff, A 1993Selective inhibition of farnesyl-protein transferase blocks ras processing in vivoJ. Biol. Chem.26876177620PubMedGoogle Scholar
  18. Giner, J.L 1998New and efficient synthetic routes to 1-Deoxy-D-xyluloseTetrahedron Lett.3924792482Google Scholar
  19. Glomset, J.A., Gelb, M.H., Farnsworth, C.C 1990Prenyl proteins in eucaryotic cells: a new type of membrane anchorTrends Biochem. Sci.15139142PubMedGoogle Scholar
  20. Hara, M., Akasaka, K., Akinaga, S., Okabe, M., Nakano, H., Gomez, R., Wood, D., Uh, M., Tamanoi, F 1993Identification of Ras farnesyltransferase inhibitors by microbial screeningProc. Natl. Acad. Sci. USA9022812285PubMedGoogle Scholar
  21. Haralampidis, K., Bryan, G., Qi, X., Papadopoulou, K., Bakht, S., Melton, R., Osbourn, A 2001A new class of oxidosqualene cyclases directs synthesis of antimicrobial phytoprotectants in monocotsProc. Natl. Acad. Sci. USA981343113436PubMedGoogle Scholar
  22. Imbault, N., Thiersault, M, Dupéron, P., Benabdelmouna, A., Doireau, P 1996Pravastatine: a tool for investigating the availability of mevalonate metabolites for primary and secondary metabolism in Catharanthus roseus cell suspensionsPhysiol. Plant98803809Google Scholar
  23. Irmler, S, Schroder, G., St-Pierre, B., Crouch, N.P., Hotze, M., Schmidt, J., Strack, D., Matern, U., Schroder, J 2000Indole alkaloid biosynthesis in Catharanthus roseus: new enzyme activities and identification of cytochrome P450 CYP72A1 as secologanin synthasePlant J.24797804PubMedGoogle Scholar
  24. Iwasaki, Y., Fujisawa, Y., Kato, H 2003Function of Heterotrimeric G Protein in Gibberellin SignallingJ. Plant Growth Regul.22126133Google Scholar
  25. Kasahara, H., Hanada, A., Kuzuyama, T., Takagi, M., Kamiya, Y., Yamaguchi, S 2002Contribution of the mevalonate and methylerythritol phosphate pathways to the biosynthesis of gibberellins in ArabidopsisJ. Biol. Chem.2774518845194PubMedGoogle Scholar
  26. Kessler, A., Baldwin, I.T 2001Defensive function of herbivore-induced plant volatile emissions in natureScience29121412144PubMedGoogle Scholar
  27. Levêque, D., Wihlm, J., Jehl, F 1996Pharmacology of Catharanthus alkaloidsBull. Cancer83176186PubMedGoogle Scholar
  28. Lichtenthaler, H.K., Schwender, J., Disch, A., Rhomer, M 1997Biosynthesis of isoprenoids in higher plant chloroplasts proceeds via a so far unexpected novel pathwayFEBS Lett.400271274PubMedGoogle Scholar
  29. Lichtenthaler, H.K 1999The 1-deoxy-D-xylulose 5-phosphate pathway of isoprenoid biosynthesis in plantsAnnu. Rev. Plant Physiol. Plant Mol. Biol.504765PubMedGoogle Scholar
  30. Mérillon, J.M., Ouelhazi, I., Doireau, P., Guillot, A., Chenieux, J.C., Rideau, M 1986Indol alkaloid accumulation and tryptophan decarboxylase activity in Catharanthus roseus cells cultured in three different mediaPlant Cell Rep.52326Google Scholar
  31. Naudascher, F., Doireau, P., Guillot, A., Viel, C., Thiersault, M 1989Time-course studies on the use of secologanin by Catharanthus roseus cells cultured in vitroJ. Plant Physiol.134608612Google Scholar
  32. Pandey, S., Assmann, S.M 2004The Arabidopsis putative G protein-coupled receptor GCR1 interacts with the G protein α subunit GPA1 and regulates abscisic acid signallingPlant Cell1616161632PubMedGoogle Scholar
  33. Papon N., Bremer J., Vansiri A., Andreu F., Rideau M. and Crèche J. Cytokinin and ethylene control indole alkaloid production at the level of the MEP/terpenoid pathway in Catharanthus roseus suspension cells. Planta Med. In pressGoogle Scholar
  34. Papon, N., Vansiri, A., Gantet, P., Chénieux, J.C., Rideau, M., Crèche, J 2004Histidine-containing phosphotransfer domain extinction by RNA interference turns off a cytokinin signalling circuitry in Catharanthus roseus suspension cellsFEBS Lett.5588588PubMedGoogle Scholar
  35. Pei, Z.M., Ghassemian, M., Kwak, C.M., Mc Court, P., Schoerder, J.I 1998Role of farnesyltransferase in ABA regulation of guard cell anion channels and plant water lossScience282287290PubMedGoogle Scholar
  36. Qian, D., Zhou, G., Ju, R., Cramer, C.L., Yang, Z 1996Protein farnesyltransferase in plants: molecular characterization and involvement in cell cycle controlPlant Cell823812394PubMedGoogle Scholar
  37. Ren, Z., Elson, C.E., Gould, M.N 1997Inhibition of type I and type II geranylgeranyl-protein transferases by the monoterpene perillyl alcohol in NIH3T3 cellsBiochem. Pharmacol.54113120PubMedGoogle Scholar
  38. Ren, Z., Gould, M.N 1994Inhibition of ubiquinone and cholesterol synthesis by the monoterpene perillyl alcoholCancer Lett.7618590PubMedGoogle Scholar
  39. Repko, E.M., Maltese, W.A 1989Post-translational isoprenylation of cellular proteins is altered in response to mevalonate availabilityJ. Biol. Chem.26499459952PubMedGoogle Scholar
  40. Rodriguez-Concepcion, M., Yalovsky, S., Zik, M., Fromm, H., Gruissem, W 1999The prenylation status of a novel plant calmodulin directs plasma membrane or nuclear localization of the proteinEMBO J.1819962007PubMedGoogle Scholar
  41. Rodriguez-Concepcion, M., Toledo-Ortiz, G., Yalovsky, S., Caldelari, D., Gruissem, W 2000Carboxyl-methylation of prenylated calmodulin CaM53 is required for efficient plasma membrane targeting of the proteinPlant J.24775784PubMedGoogle Scholar
  42. Running, M.P., Lavy, M., Sternberg, H., Galichet, A., Gruissem, W., Hake, S., Ori, N., Yalosky, S 2004Enlarged meristems and delayed growth in plp mutants result from lack of CaaX prenyltransferasesProc. Natl. Acad. Sci. USA10178157820PubMedGoogle Scholar
  43. Sambrook, J., Fritsch, E.F., Maniatis, T. 1989Molecular Cloning: A Laboratory Manual, Ed 2Cold Spring Harbor Laboratory PressCold Spring Harbor, NYGoogle Scholar
  44. St-Pierre, B., Vazquez-Flota, F.A., De Luca, V 1999Multicellular compartimentation of Catharanthus roseus alkaloid biosynthesis predicts intercellular translocation of a pathway intermediatePlant Cell11887900PubMedGoogle Scholar
  45. Tamanoi, F 1993Inhibitors of Ras farnesyltransferasesTrends Biochem Sci8349353Google Scholar
  46. Tao, L.Z., Cheung, A.Y., Wu, H.M 2002Plant Rac-Like GTPases are activated by auxin and mediate auxin-responsive gene expressionPlant Cell1427452760PubMedGoogle Scholar
  47. Ullah, H., Chen, J.G., Temple, B., Boyes, D.C., Alonso, J.M., Davis, K.R., Ecker, J.R., Jones, A.M 2003The beta-subunit of the Arabidopsis G protein negatively regulates auxin-induced cell division and affects multiple developmental processesPlant Cell15393409PubMedGoogle Scholar
  48. Van Telligen, O., Sips, J.H.M., Beijnen, J.H., Bult, A., Nooijen, W.J 1992Pharmacology, bio-analysis and pharmacokinetics in the Vinca alkaloids and semi-synthetic derivativesAnticancer Res.1216991716PubMedGoogle Scholar
  49. Veau, B., Courtois, M., Oudin, A., Chénieux, J.C., Rideau, M., Clastre, M 2000Cloning and expression of cDNAs encoding two enzymes of the MEP pathway in Catharanthus roseusBiochim. Biophys. Acta1517159163PubMedGoogle Scholar
  50. Walter, M.H., Hans, J., Strack, D 2002Two distantly related genes encoding 1-deoxy-d-xylulose 5-phosphate synthases: differential regulation in shoots and apocarotenoid-accumulating mycorrhizal rootsPlant J.31243254PubMedGoogle Scholar
  51. Wesley, S.V., Helliwell, C.A., Smith, N.A., Wang, M.B., Rouse, D.T., Liu, Q., Gooding, P.S., Singh, S.P., Abbott, D., Stoutjesdijk, P.A., Robinson, S.P., Gleave, A.P., Green, A.G., Waterhouse, P.M 2001Construct design for efficient, effective and high-throughput gene silencing in plantsPlant J.27581590Google Scholar
  52. Wink M. 1999. Introduction: biochemistry, role and biotechnology of secondary metabolites. In: M. Wink (Ed), Biochemistry of plant secondary metabolism Vol. 2, Sheffield Academic Press and CRC Press, Annual Plant Reviews, pp. 1–16Google Scholar
  53. Yalovsky, S., Rodriguez-Concepcion, M., Bracha, K., Toledo-Ortiz, G., Gruissem, W 2000aPrenylation of the floral transcription factor APETALA1 modulates its functionPlant Cell1212571266Google Scholar
  54. Yalovsky, S., Kulukiam, A., Rodriguez-Concepcion, M., Young, C.A., Gruissem, W 2000bFunctional requirement of plant farnesyltransferase during development in ArabidopsisPlant Cell1212671278Google Scholar
  55. Yang, Y., Shah, J., Klessing, D.F 1997Signal perception and transduction in plant defense responsesGenes Dev.1116211639PubMedGoogle Scholar
  56. Yang, Z., Cramer, C.L., Watson, J. 1993Protein farnesyltransferase in plants. Molecular cloning and expression of a homolog of the beta subunit from the garden PeaPlant Physiol.101667674PubMedGoogle Scholar
  57. Zheng, Z.L., Yang, Z 2000The Rop GTPase: an emerging signalling switch in plantsPlant Mol. Biol.4419PubMedGoogle Scholar
  58. Zhang, F.L., Casey, P.J 1996Protein prenylation: molecular mechanisms and functional consequencesAnnu. Rev. Biochem.65241269PubMedGoogle Scholar
  59. Ziegelhoffer, E.C., Medrano, L.J., Meyerowitz, E.M 2000Cloning of the Arabidopsis WIGGUM gene identifies a role for farnesylation in meristem developmentProc. Natl. Acad. Sci. USA9776337638PubMedGoogle Scholar

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

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