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

Overview

Journal F1000Res

Specialties Biomedical Engineering
Science

Date 2014 Feb 25

PMID 24555083

Citations 5

Authors

Michael Hartmann

Andrea Hemmerlin

Elisabet Gas-Pascual

Esther Gerber

Denis Tritsch

Michel Rohmer

Thomas J Bach

Affiliations

Soon will be listed here.

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.

Citing Articles

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Fosmidomycin biosynthesis diverges from related phosphonate natural products.

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Development of an image-based screening system for inhibitors of the plastidial MEP pathway and of protein geranylgeranylation.

Hartmann M, Gas-Pascual E, Hemmerlin A, Rohmer M, Bach T F1000Res. 2017; 4:14.

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Past achievements, current status and future perspectives of studies on 3-hydroxy-3-methylglutaryl-CoA synthase (HMGS) in the mevalonate (MVA) pathway.

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