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Title: The plastidial 2-C-methyl-D-erythritol 4-phosphate pathway provides the isoprenyl moiety for protein geranylgeranylation in tobacco BY-2 cells .
Author: Gerber E Hemmerlin A Hartmann M Heintz D Hartmann MA Mutterer J Rodriguez-Concepcion M Boronat A Van Dorsselaer A Rohmer M Crowell DN Bach TJ
Journal: Plant Cell Citation: V : 21 P : 285-300 Year: 2009 Type: MEDLINE
Literature: oryza Field: abstract Doc ID: pub19136647 Accession (PMID): 19136647
Abstract: Protein farnesylation and geranylgeranylation are important posttranslational modifications in eukaryotic cells . We visualized in transformed Nicotiana tabacum Bright Yellow-2 ( BY-2 ) cells the geranylgeranylation and plasma membrane localization of GFP-BD-CVIL , which consists of green fluorescent protein ( GFP ) fused to the C-terminal polybasic domain ( BD ) and CVIL isoprenylation motif from the Oryza sativa calmodulin , CaM61 . Treatment with fosmidomycin ( Fos ) or oxoclomazone ( OC ) , inhibitors of the plastidial 2-C-methyl-d-erythritol 4-phosphate ( MEP ) pathway , caused mislocalization of the protein to the nucleus , whereas treatment with mevinolin , an inhibitor of the cytosolic mevalonate pathway , did not . The nuclear localization of GFP-BD-CVIL in the presence of MEP pathway inhibitors was completely reversed by all-trans-geranylgeraniol ( GGol ) . Furthermore , 1-deoxy-d-xylulose ( DX ) reversed the effects of OC , but not Fos , consistent with the hypothesis that OC blocks 1-deoxy-d-xylulose 5-phosphate synthesis , whereas Fos inhibits its conversion to 2-C-methyl-d-erythritol 4-phosphate . By contrast , GGol and DX did not rescue the nuclear mislocalization of GFP-BD-CVIL in the presence of a protein geranylgeranyltransferase type 1 inhibitor . Thus , the MEP pathway has an essential role in geranylgeranyl diphosphate ( GGPP ) biosynthesis and protein geranylgeranylation in BY-2 cells . GFP-BD-CVIL is a versatile tool for identifying pharmaceuticals and herbicides that interfere either with GGPP biosynthesis or with protein geranylgeranylation .
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[ Sen. 2, subscore: 1.00 ]: We visualized in transformed Nicotiana tabacum Bright Yellow-2 ( BY-2 ) cells the geranylgeranylation and plasma membrane localization of GFP-BD-CVIL , which consists of green fluorescent protein ( GFP ) fused to the C-terminal polybasic domain ( BD ) and CVIL isoprenylation motif from the Oryza sativa calmodulin , CaM61 .
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