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Geranylgeranyl pyrophosphate synthase encoded by the newly isolated gene GGPS6 from Arabidopsis thaliana is localized in mitochondria
Authors:Zhu  XuFen  Suzuki  Kengo  Saito  Takeshi  Okada  Kazunori  Tanaka  Katsunori  Nakagawa  Tsuyoshi  Matsuda  Hideyuki  Kawamukai  Makoto
Institution:(1) Department of Applied Bioscience and Biotechnology, Faculty of Life and Environmental Science, Japan;(2) Research Institute of Molecular Genetics, Shimane University, 1060 Nishikawatsu, Matsue, 690, Japan
Abstract:We have cloned a new geranylgeranyl pyrophosphate (GGPP) synthase gene, designated GGPS6/, from Arabidopsis thaliana genomic DNA. Nucleotide sequence analysis revealed that the GGPS6 gene contains an open reading frame coding for a protein of 343 amino acid residues with a calculated molecular mass of 37 507 Da. Also, the gene is not interrupted by an intron. The predicted amino acid sequence of the GGPS6 gene shows significant homology (34.0–57.7%) with other GGPP synthases from Arabidopsis. The GGPS6 protein contains a N-terminal signal peptide which is thought to function as an organelle targeting sequence. In fact, the GGPS6-GFP fusion protein was found to be localized exclusively in mitochondria when expressed in tobacco BY-2 cells. In vitro analysis of the enzyme activity as well as genetic complementation analysis with Erwinia uredovora crt gene cluster expressed in Escherichia coli showed that the GGPS6 gene most certainly encodes a GGPP synthase catalyzing the conversion of farnesyl pyrophosphate to GGPP.
Keywords:Arabidopsis thaliana/  carotenoid  crtE/  gene cloning  GGPP synthase  mitochondria
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