Okada K, Saito T, Nakagawa T, Kawamukai M, Kamiya Y
Frontier Research Program Plant Hormone Function, The Institute of Physical and Chemical Research (RIKEN), Hirosawa 2-1, Wako-shi, Saitama, 351-0198, Japan.
Plant Physiol. 2000 Apr;122(4):1045-56. doi: 10.1104/pp.122.4.1045.
Geranylgeranyl diphosphate (GGPP) is the precursor for the biosynthesis of gibberellins, carotenoids, chlorophylls, isoprenoid quinones, and geranylgeranylated proteins in plants. There is a small gene family for GGPP synthases encoding five isozymes and one related protein in Arabidopsis, and all homologs have a putative localization signal to translocate into specific subcellular compartments. Using a synthetic green fluorescent protein (sGFP), we studied the subcellular localization of these GGPP synthases. When these fusion proteins were expressed by the cauliflower mosaic virus 35S promoter in Arabidopsis, GGPS1-sGFP and GGPS3-sGFP proteins were translocated into the chloroplast, GGPS2-sGFP and GGPS4-sGFP proteins were localized in the endoplasmic reticulum, and the GGPS6-sGFP protein was localized in the mitochondria. Both GGPS1 and GGPS3 proteins synthesized in vitro were taken up into isolated intact pea chloroplasts and processed to the mature form. RNA-blot and promoter-beta-glucuronidase (GUS) analysis showed that these GGPP synthases genes are organ-specifically expressed in Arabidopsis. GGR and GGPS1 were ubiquitously expressed, while GGPS2, GGPS3, and GGPS4 were expressed specifically in the flower, root, and flower, respectively. These results suggest that each GGPP synthase gene is expressed in different tissues during plant development and GGPP is synthesized by the organelles themselves rather than being transported into the organelles. Therefore, we predict there will be specific pathways of GGPP production in each organelle.
香叶基香叶基二磷酸(GGPP)是植物中赤霉素、类胡萝卜素、叶绿素、类异戊二烯醌和香叶基香叶基化蛋白生物合成的前体。在拟南芥中,编码五种同工酶和一种相关蛋白的GGPP合酶存在一个小基因家族,所有同源物都有一个推定的定位信号,可转运到特定的亚细胞区室。利用合成绿色荧光蛋白(sGFP),我们研究了这些GGPP合酶的亚细胞定位。当这些融合蛋白由花椰菜花叶病毒35S启动子在拟南芥中表达时,GGPS1-sGFP和GGPS3-sGFP蛋白转运到叶绿体中,GGPS2-sGFP和GGPS4-sGFP蛋白定位于内质网中,而GGPS6-sGFP蛋白定位于线粒体中。体外合成的GGPS1和GGPS3蛋白都被摄取到分离的完整豌豆叶绿体中,并加工成成熟形式。RNA印迹和启动子-β-葡萄糖醛酸酶(GUS)分析表明,这些GGPP合酶基因在拟南芥中是器官特异性表达的。GGR和GGPS1普遍表达,而GGPS2、GGPS3和GGPS4分别在花、根和花中特异性表达。这些结果表明,每个GGPP合酶基因在植物发育过程中的不同组织中表达,并且GGPP是由细胞器自身合成的,而不是被转运到细胞器中。因此,我们预测每个细胞器中会有特定的GGPP产生途径。