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PMID: 15604701 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

The AtPPT1 gene encoding 4-hydroxybenzoate polyprenyl diphosphate transferase in ubiquinone biosynthesis is required for embryo development in Arabidopsis thaliana.

Plant molecular biology ·Vol. 55 ·No. 4 ·2004-07-00 ·Pages 567-77

Okada K, Ohara K, Yazaki K, Nozaki K, Uchida N, Kawamukai M, Nojiri H, Yamane H

Abstract

4-Hydroxybenzoate polyprenyl diphosphate transferase (4HPT) is the key enzyme that transfers the prenyl side chain to the benzoquione frame in ubiquinone (UQ) biosynthesis. The Arabidopsis AtPPT1 cDNA encoding 4HPT was cloned by reverse transcription-polymerase chain reaction (RT-PCR) based on the information of the Arabidopsis genomic sequence, and the function of the gene was determined. Heterologous expression of the AtPPT1 gene enabled restoration of the respiratory ability and UQ synthesis in a yeast mutant that was defective in 4HPT activity. The mitochondrial fraction that was prepared from the yeast mutant, which expressed the AtPPT1 gene, exhibited 4HPT enzymatic activity with geranyl diphosphate (GPP) as the prenyl substrate. This indicated that the AtPPT1 gene encodes active 4HPT with a broad substrate specificity in terms of the prenyl donor. The AtPPT1 mRNA was predominantly expressed in the flower cluster, and the green fluorescent protein (GFP) fused with the signal peptide of AtPPT1 was translocated into the mitochondria. T-DNA insertion mutation that disrupts the AtPPT1 gene in Arabidopsis resulted in the arrest of embryo development at an early stage of zygotic embryogenesis. These results demonstrate that the AtPPT1 gene involved in the biosynthesis of mitochondrial UQ plays an essential role in embryo development in Arabidopsis .

MeSH Terms
Alkyl and Aryl Transferases/genetics,metabolism Amino Acid Sequence Arabidopsis/embryology,genetics,metabolism Blotting, Northern Cloning, Molecular DNA, Complementary/chemistry,genetics Gene Expression Regulation, Developmental Gene Expression Regulation, Enzymologic Gene Expression Regulation, Plant Genetic Complementation Test Green Fluorescent Proteins/genetics,metabolism Microscopy, Confocal Molecular Sequence Data Mutation RNA, Messenger/genetics,metabolism Recombinant Fusion Proteins/genetics,metabolism Reverse Transcriptase Polymerase Chain Reaction Saccharomyces cerevisiae/genetics Seeds/enzymology,genetics,growth & development Sequence Alignment Sequence Analysis, DNA Sequence Homology, Amino Acid Substrate Specificity Ubiquinone/biosynthesis
Chemicals
DNA, Complementary RNA, Messenger Recombinant Fusion Proteins Ubiquinone Green Fluorescent Proteins Alkyl and Aryl Transferases 4-hydroxybenzoate polyprenyltransferase
Authors & Affiliations
8 authors, click to expand affiliations / ORCID
Okada Kazunori
Laboratory of Environmental Biochemistry, Biotechnology Research Center, The University of Tokyo, 1-1-1 Yayoi, Tokyo, Japan. ukokada@mail.ecc.u-tokyo.ac.jp
Ohara Kazuaki
Yazaki Kazufumi
Nozaki Kouhei
Uchida Naonori
Kawamukai Makoto
Nojiri Hideaki
Yamane Hisakazu
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Article Info
Journal
Plant molecular biology
Abbr.
Plant Mol Biol
ISSN
0167-4412
Published
2004-07-00
Pages
567-77
Language
English
Region
Netherlands
NLM ID
9106343
Subset
IM
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