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 [v2; ref status: indexed, http://f1000r.es/2af]
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 g...
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F1000 Research Ltd
2013-11-01
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| Online Access: | http://f1000research.com/articles/2-170/v2 |
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| author | Michael Hartmann Andrea Hemmerlin Elisabet Gas-Pascual Esther Gerber Denis Tritsch Michel Rohmer Thomas J Bach |
| author_facet | Michael Hartmann Andrea Hemmerlin Elisabet Gas-Pascual Esther Gerber Denis Tritsch Michel Rohmer Thomas J Bach |
| author_sort | Michael Hartmann |
| collection | DOAJ |
| description | 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. |
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| institution | Directory Open Access Journal |
| issn | 2046-1402 |
| language | English |
| last_indexed | 2024-12-10T21:58:50Z |
| publishDate | 2013-11-01 |
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| spelling | doaj.art-e6058733c12c478bb9cd54fb3d9ba3d72022-12-22T01:31:57ZengF1000 Research LtdF1000Research2046-14022013-11-01210.12688/f1000research.2-170.v22967The 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 [v2; ref status: indexed, http://f1000r.es/2af]Michael Hartmann0Andrea Hemmerlin1Elisabet Gas-Pascual2Esther Gerber3Denis Tritsch4Michel Rohmer5Thomas J Bach6Current address: Institute of Biological Chemistry, Washington State University, Pullman WA, 99164-6340, USADépartement Réseaux Métaboliques, Institut de Biologie Moléculaire des Plantes, CNRS UPR 2357, Université de Strasbourg, F-67083 Strasbourg, FranceCurrent address: Department of Horticulture and Crop Science, Ohio State University, Wooster OH, 44691, USACurrent address: Deinove SA, F-34830 Clapiers, FranceUMR 7177 CNRS, Laboratoire de Chimie et Biochimie des Microorganismes, Institut de Chimie de Strasbourg, Université de Strasbourg, F-67008 Strasbourg, FranceUMR 7177 CNRS, Laboratoire de Chimie et Biochimie des Microorganismes, Institut de Chimie de Strasbourg, Université de Strasbourg, F-67008 Strasbourg, FranceDépartement Réseaux Métaboliques, Institut de Biologie Moléculaire des Plantes, CNRS UPR 2357, Université de Strasbourg, F-67083 Strasbourg, FranceWe 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.http://f1000research.com/articles/2-170/v2Agriculture & BiotechnologyDrug Discovery & DesignPlant Biochemistry & PhysiologyPlant Cell Biology |
| spellingShingle | Michael Hartmann Andrea Hemmerlin Elisabet Gas-Pascual Esther Gerber Denis Tritsch Michel Rohmer Thomas J Bach 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 [v2; ref status: indexed, http://f1000r.es/2af] F1000Research Agriculture & Biotechnology Drug Discovery & Design Plant Biochemistry & Physiology Plant Cell Biology |
| 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 [v2; ref status: indexed, http://f1000r.es/2af] |
| title_full | 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 [v2; ref status: indexed, http://f1000r.es/2af] |
| title_fullStr | 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 [v2; ref status: indexed, http://f1000r.es/2af] |
| title_full_unstemmed | 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 [v2; ref status: indexed, http://f1000r.es/2af] |
| title_short | 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 [v2; ref status: indexed, http://f1000r.es/2af] |
| title_sort | 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 v2 ref status indexed http f1000r es 2af |
| topic | Agriculture & Biotechnology Drug Discovery & Design Plant Biochemistry & Physiology Plant Cell Biology |
| url | http://f1000research.com/articles/2-170/v2 |
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