if the gene encoding a specific enzyme in the plastoquinone

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22 January 2021

This stop codon results in the deletion of the remaining 26 amino acids from the carboxyterminal end of the protein. The first step of this pathway, common to the synthesis of both plastoquinone and tocopherol, is the formation of HGA from HPP by the enzyme HPPDase (EC 1.13.11.27). The role of metabolism in the antioxidant function of vitamin E. Treatment of hereditary tyrosinemia type I by inhibition of 4 hydroxyphenylpyruvate dioxygenase. Plastoquinone-deficient mutants of maize and A. thaliana exhibit severe growth defects and seedling lethality (7, 9, 32). Similar results were reported when aS. Similarly, for the pds1 mutant, two sets of primers were used: SN418T7+10 and SN418MF+1b (5′-CAGATGTTGTAGCCCT-3′) for the first 1000 bp of the gene, and SN418T7+4 and SN418MF+12 for the last 700 bp of the gene. 6/f complex, and to a lesser extent as an electron carrier for NAD(P)H-plastoquinone oxidoreductases (Berger et al., 1993). 2). ↵1 Present address: Boyce Thompson Institute, Tower Road, Ithaca, NY 14853. ScienceDirect ® is a registered trademark of Elsevier B.V. ScienceDirect ® is a registered trademark of Elsevier B.V. For complementation analysis, kanamycin-resistant T2plants were crossed with PDS1/pds1 heterozygotes. The putative Arabidopsis HPPDase protein has from 17% to 27% amino acid identity with bacterial, fungal, and animal HPPDases and between 58% and 70% amino acid identity with two other plant HPPDases. The 460-bp insert from this expressed sequence tag was used as a probe to screen 4 × 105 plaques of the Arabidopsis PRL2 library (Newman et al., 1994). They are also α-tocopherol … AJ000693, D64004, L38493, U11864, U87257, S69666,M59289, M59429, Z50016, X72389, D29987, M18405, and D13390). Kanamycin-resistant F1 plants were selected and selfed, and the resulting F2 plants were scored. Both HPPDase genomic sequences contain a single 107-bp intron of identical sequence between positions 1162 and 1163 of the HPPD cDNA sequence in Figure 2. Mutation of the PDS1 locus disrupts the activity of p-hydroxyphenylpyruvate dioxygenase (HPPDase), the first committed step in the synthesis of both plastoquinone and tocopherols in plants. A versatile binary vector system with a T-DNA organisational structure conducive to efficient integration of cloned DNA into the plant genome. Genes and cDNAs encoding HPPDase have been identified from several mammalian, fungal, bacterial, and plant sources (Gershwin et al., 1987;Endo et al., 1992, 1995; Hummel et al., 1992; Ruetschi et al., 1993;Coon et al., 1994; Denoya et al., 1994; Wilson et al., 1994;Wintermeyer et al., 1994; Kaneko et al., 1995; Wyckoff et al., 1995;Garcia et al., 1997) and show between 25% and 95% identity at the amino acid level. Their mode of action arises from a direct inhibition of plastoquinone and tocopherol synthesis and an indirect inhibition of carotenoid desaturation (Mayonado et al., 1989;Schultz et al., 1993; Secor, 1994). Finally, and most significantly, we have shown that the pds1 HPPD gene contains a small deletion that results in the elimination of a portion of the carboxy terminus of the protein. The occurrence of multiple genes encoding HMGR is a general feature of higher plants: two genes are present in Arabidopsis (Caelles et al., 1989; Enjuto et al., 1994), three genes have been reported in Hevea brasiliensis (Chye et al., 1992), and four genes are present in … The middle and bottom plots are cell-free extracts from cultures of E. coliharboring the pET-HPPD construct and the pET15b construct, respectively. These data demonstrate that the Arabidopsis cDNA pHPPD encodes a functional HPPDase enzyme. The plastoquinone and α-tocopherol biosynthetic pathway in higher plants. Presumably, these highly conserved residues are important for substrate binding or the catalytic mechanism of HPPDases. In this paper, we report the cloning and functional analysis of gene products from Synechocystis sp. A similar dark-brown coloration was reported when the gene encoding HPPDase fromStreptomyces avermitilis was expressed in E. coli(Denoya et al., 1994). Plastoquinone and tocopherols are the two major classes of chloroplastic, lipid-soluble quinone compounds in higher plants. As a result of the central role HPPDase serves in aromatic amino acid metabolism in mammals and plastidic quinone synthesis in plants, a class of competitive inhibitors of HPPDases collectively known as triketones has been developed and used for a variety of clinical and agricultural purposes (Lindstedt et al., 1992; Schultz et al., 1993; Secor, 1994). FQR is a putative enzyme that reduces plastoquinone using Fd as a direct electron donor and has been characterized as a binding site for antimycin A, a specific inhibitor of cyclic electron flow. Genes galore: a summary of methods for accessing results from large-scale partial sequencing of anonymous Arabidopsis cDNA clones. This paper reports the isolation of a cDNA, pHPPD, encoding Arabidopsis HPPDase and its functional characterization by expression in both plants and Escherichia coli. Arabidopsis HPPDase amino acid residues showing identity in 9 of the other 13 HPPDase proteins are indicated with shaded boxes. Five Tyr and His residues, postulated to form a ferric iron center in HPPDases (Denoya et al., 1994), are also conserved in the putative Arabidopsis HPPDase (Fig. As shown in Figure 3, E. coli cultures expressing pHPPD accumulate a compound that co-migrates with, and has a spectrum identical to, the HGA standard (Fig. AF060481) and pds1 mutant tissues were determined by direct sequencing of PCR-amplified products. The pET15b control culture lacked a peak at 7.9 min and had a minor peak at 7.7 min, with a spectrum and absorbance maxima (271, 280, and 287 nm) that indicated that it was not HGA (Fig.3B). Our results indicate that in Synechocystis in contrast to the situation in higher plants the 4-hydroxyphenylpyruvate dioxygenase is not required for the synthesis of plastoquinone. In previous studies we characterized two loci in Arabidopsis defining key steps of this biosynthetic pathway. Peripheral neuropathy as the presenting feature of tyrosinaemia type I and effectively treated with an inhibitor of 4-hydroxyphenylpyruvate dioxygenase. In this study, we isolated and identified 10 MEP pathway genes in the important aromatic plant sweet osmanthus (Osmanthus fragrans). In mammals, which cannot synthesize plastoquinone or tocopherols, α-tocopherol (vitamin E) is an essential dietary component (Mason, 1980) and has a well-documented role as a membrane-associated free radical scavenger (for review, seeLiebler, 1993). Combined, these data conclusively demonstrate that pds1 is a mutation of the HPPDase gene. T20952) is indicated by a single underline. In previous work we demonstrated that the biochemical basis of the Arabidopsis pds1 mutation is an inability to convert HPP to HGA (Fig. Fifty percent of the resulting kanamycin-resistant F1 progeny from these crosses were also heterozygous for the pds1 mutation. Genomic DNAs for use as substrates for PCR were isolated from wild-type and pds1 genotypes (both are ecotype Wassilewskija [Ws]) by the modified minipreparation method (DellaPorta et al., 1983). This question is for testing whether or not you are a human visitor and to prevent automated spam submissions. 1). Evaluation of the mechanism of action of the bleaching herbicide SC-0051 by HPLC analysis. Amino acid residues identical in all 14 HPPDase sequences are denoted with black boxes. The first step of this pathway, common to the synthesis of both plastoquinone and tocopherol, is the formation of HGA from HPP by the enzyme HPPDase (EC 1.13.11.27). of chlorophyll biosynthesis enzymes and that of the Lhcb genes in the model organism Dunaliella salina. The locations of the pds1 andpds2 mutations in the pathway are indicated. In plants, triketones such as sulcotrione (2-[4-chloro-2-nitrobenzoyl]-5,5-dimethylcyclohexane-1,3-dione) are effective bleaching herbicides. NOTE: We only request your email address so that the person you are recommending the page to knows that you wanted them to see it, and that it is not junk mail. As shown in Figure 2, the wild-type and pds1 HPPD alleles are identical in sequence with the exception of a 17-bp deletion in the pds1HPPD allele. Table I shows that the F2 green-to-white segregation ratios from crosses of pds1 heterozygotes to three independent, parental transgenic lines are statistically significant for a 15:1 ratio. HGA was identified in extracts based on comparison of retention time and spectra to a HGA (Sigma) standard with a Hewlett-Packard series 1100 chromatograph and photodiode array detector. Plastoquinone-9 (PQ-9) is an essential component of photosynthesis that carries electrons in the linear and alternative electron transport chains, and is also a redox sensor that regulates state transitions and gene expression. Eleven genes encoding chloroplast NADH dehydrogenase-like (NDH) complex have been discovered in plastid genomes on the basis of their homology with genes encoding respiratory complex I. Of higher plant chloroplasts and are ubiquitous in plants, especially in mitochondria and chloroplast constitutively HPPDase! Light energy and to prevent automated spam submissions activity is often present at very high levels is!, tyrosine-derived metabolites, tocopherols, they do nonetheless contain HPPDase enzymatic.... 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