title: The effect of MEP pathway and other inhibitors on the intracellular localization of a plasma membrane-targeted, isoprenylable GFP reporter protein in tobacco BY-2 cells.
abstract: We have established an in vivo visualization system for the geranylgeranylation of proteins in a stably transformed tobacco BY-2 cell line, based on the expression of a dexamethasone-inducible GFP fused to the carboxy-terminal basic domain of the rice calmodulin CaM61, which naturally bears a CaaL geranylgeranylation motif (GFP-BD-CVIL). By using pathway-specific inhibitors it was demonstrated that inhibition of the methylerythritol phosphate (MEP) pathway with known inhibitors like oxoclomazone and fosmidomycin, as well as inhibition of the protein geranylgeranyltransferase type 1 (PGGT-1), shifted the localization of the GFP-BD-CVIL protein from the membrane to the nucleus. In contrast, the inhibition of the mevalonate (MVA) pathway with mevinolin did not affect the localization. During the present work, this test system has been used to examine the effect of newly designed inhibitors of the MEP pathway and inhibitors of sterol biosynthesis such as squalestatin, terbinafine and Ro48-8071. In addition, we also studied the impact of different post-prenylation inhibitors or those suspected to affect the transport of proteins to the plasma membrane on the localization of the geranylgeranylable fusion protein GFP-BD-CVIL.
authors.affiliation: Département Réseaux Métaboliques, Institut de Biologie Moléculaire des Plantes, CNRS UPR 2357, Université de Strasbourg, F-67083 Strasbourg, France ; Current address: Institute of Biological Chemistry, Washington State University, Pullman WA, 99164-6340, USA.
authors.forename: Michael
authors.initials: M
authors.lastname: Hartmann
authors.affiliation: Département Réseaux Métaboliques, Institut de Biologie Moléculaire des Plantes, CNRS UPR 2357, Université de Strasbourg, F-67083 Strasbourg, France.
authors.forename: Andrea
authors.initials: A
authors.lastname: Hemmerlin
authors.affiliation: Département Réseaux Métaboliques, Institut de Biologie Moléculaire des Plantes, CNRS UPR 2357, Université de Strasbourg, F-67083 Strasbourg, France ; Current address: Department of Horticulture and Crop Science, Ohio State University, Wooster OH, 44691, USA.
authors.forename: Elisabet
authors.initials: E
authors.lastname: Gas-Pascual
authors.affiliation: Département Réseaux Métaboliques, Institut de Biologie Moléculaire des Plantes, CNRS UPR 2357, Université de Strasbourg, F-67083 Strasbourg, France ; Current address: Deinove SA, F-34830 Clapiers, France.
authors.forename: Esther
authors.initials: E
authors.lastname: Gerber
authors.affiliation: UMR 7177 CNRS, Laboratoire de Chimie et Biochimie des Microorganismes, Institut de Chimie de Strasbourg, Université de Strasbourg, F-67008 Strasbourg, France.
authors.forename: Denis
authors.initials: D
authors.lastname: Tritsch
authors.affiliation: UMR 7177 CNRS, Laboratoire de Chimie et Biochimie des Microorganismes, Institut de Chimie de Strasbourg, Université de Strasbourg, F-67008 Strasbourg, France.
authors.forename: Michel
authors.initials: M
authors.lastname: Rohmer
authors.affiliation: Département Réseaux Métaboliques, Institut de Biologie Moléculaire des Plantes, CNRS UPR 2357, Université de Strasbourg, F-67083 Strasbourg, France.
authors.forename: Thomas J
authors.initials: TJ
authors.lastname: Bach