Cloning of the sdsA gene encoding solanesyl diphosphate synthase from Rhodobacter capsulatus and its functional expression in Escherichia coli and Saccharomyces cerevisiae

Okada, Kazunori; Kamiya, Yasuhiro; Zhu, Xiaoli; Suzuki, Kengo; Tanaka, Katsunori; Nakagawa, Tsuyoshi; Matsuda, Hideyuki; Kawamukai, Makoto

doi:10.1128/jb.179.19.5992-5998.1997

Cited by 62 publications

(49 citation statements)

References 30 publications

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“…We succeeded in complementing E. coli deficient in polyprenyl-diphosphate synthase. A number of genes encoding polyprenyl-diphosphate synthases from bacteria have been successfully expressed in these E. coli mutants (33,60). The TcSPPS protein is assumed to work as a homodimer, because it is functional without any other genes, unlike the case of Bacillus subtilis heptaprenyl-diphosphate synthase (heterodimer) (61), fission yeast decaprenyl-diphosphate synthase (62), mouse solanesyl-diphosphate synthase (63), and human decaprenyldiphosphate synthase (heterotetramers) (63).…”

Section: Discussionmentioning

confidence: 99%

A Solanesyl-diphosphate Synthase Localizes in Glycosomes of Trypanosoma cruzi

Ferella

Montalvetti

Rohloff

et al. 2006

Journal of Biological Chemistry

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We report the cloning of a Trypanosoma cruzi gene encoding a solanesyl-diphosphate synthase, TcSPPS. The amino acid sequence (molecular mass ϳ 39 kDa) is homologous to polyprenyl-diphosphate synthases from different organisms, showing the seven conserved motifs and the typical hydrophobic profile. TcSPPS preferred geranylgeranyl diphosphate as the allylic substrate. The final product, as determined by TLC, had nine isoprene units. This suggests that the parasite synthesizes mainly ubiquinone-9 (UQ-9), as described for Trypanosoma brucei and Leishmania major. In fact, that was the length of the ubiquinone extracted from epimastigotes, as determined by high-performance liquid chromatography. Expression of TcSPPS was able to complement an Escherichia coli ispB mutant. A punctuated pattern in the cytoplasm of the parasite was detected by immunofluorescence analysis with a specific polyclonal antibody against TcSPPS. An overlapping fluorescence pattern was observed using an antibody directed against the glycosomal marker pyruvate phosphate dikinase, suggesting that this step of the isoprenoid biosynthetic pathway is located in the glycosomes. Co-localization in glycosomes was confirmed by immunogold electron microscopy and subcellular fractionation. Because UQ has a central role in energy production and in reoxidation of reduction equivalents, TcSPPS is promising as a new chemotherapeutic target.

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Section: Discussionmentioning

confidence: 99%

A Solanesyl-diphosphate Synthase Localizes in Glycosomes of Trypanosoma cruzi

Ferella

Montalvetti

Rohloff

et al. 2006

Journal of Biological Chemistry

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“…5) As for the enzymes giving these precursors, several genes isolated from microorganisms have been well characterized, [6][7][8] but information about the plant-origin enzymes has been limited. Recently, we succeeded in the molecular cloning of Arabidopsis thaliana solanesyl diphosphate (SPP; C 45 ) synthase (At-SPS1; formerly designated At-SPS), which catalyzes the trans-type condensation of IPP to yield the C 45 product.…”

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Functional Analysis of Two Solanesyl Diphosphate Synthases fromArabidopsis thaliana

Hirooka

Izumi

et al. 2005

Bioscience, Biotechnology, and Biochemistry

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Solanesyl diphosphate (SPP) is regarded as the precursor of the side-chains of both plastoquinone and ubiquinone in Arabidopsis thaliana. We previously analyzed A. thaliana SPP synthase (At-SPS1) (Hirooka et al., Biochem. J., 370, 679-686 (2003)). In this study, we cloned a second SPP synthase (At-SPS2) gene from A. thaliana and characterized the recombinant protein.Kinetic analysis indicated that At-SPS2 prefers geranylgeranyl diphosphate to farnesyl diphosphate as the allylic substrate. Several of its features, including the substrate preference, were similar to those of At-SPS1. These data indicate that At-SPS1 and At-SPS2 share their basic catalytic machinery. Moreover, analysis of the subcellular localization by the transient expression of green fluorescent protein-fusion proteins showed that At-SPS2 is transported into chloroplasts, whereas At-SPS1 is likely to be localized in the endoplasmic reticulum in the A. thaliana cells. It is known that the ubiquinone side-chain originates from isopentenyl diphosphate derived from the cytosolic mevalonate pathway, while the plastoquinone side-chain is synthesized from isopentenyl diphosphate derived from the plastidial methylerythritol phosphate pathway. Based on this information, we propose that At-SPS1 contributes to the biosynthesis of the ubiquinone side-chain and that At-SPS2 supplies the precursor of the plastoquinone side-chain in A. thaliana.Key words: isoprenoid; prenyltransferase; nonaprenyl diphosphate; plastoquinone; ubiquinone Plants have two major prenylquinones, plastoquinone and ubiquinone. 1) Although both share the structural feature of a trans-polyprenyl tail attached to the benzoquinone skeleton and have common oxidationreduction properties, their subcellular localization and biochemical roles are distinct. Plastoquinone exists in the thylakoid membrane of the chloroplast and acts as an electron carrier in the photosynthetic electron transfer reaction,2) whereas ubiquinone exists in the inner membrane of the mitochondrion and transfers electrons in the respiratory chain reaction.3)The hydrophobic tails of prenylquinones are C 30 -C 50 in length and serve as membrane anchors. The lengths of the polyprenyl side-chains differ between plastoquinone and ubiquinone, and even among plant species.4) It is thought that these polyprenyl chains are derived from C 30 -C 50 prenyl diphosphates formed by the consecutive condensation of isopentenyl diphosphate (IPP; C 5 ) with allylic diphosphate in the trans-configuration.5) As for the enzymes giving these precursors, several genes isolated from microorganisms have been well characterized, [6][7][8] but information about the plant-origin enzymes has been limited. Recently, we succeeded in the molecular cloning of Arabidopsis thaliana solanesyl diphosphate (SPP; C 45 ) synthase (At-SPS1; formerly designated At-SPS), which catalyzes the trans-type condensation of IPP to yield the C 45 product. 9) Enzymological analysis indicated that At-SPS1 utilizes both farnesyl diphosphate (FPP; C 15 ) and geranylgeranyl di...

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“…To further understand the effects of tail length on Q function and transport, we utilized E. coli strains that had been engineered to produce quinones with different tail lengths (17,18). The polyprenyldiphosphate synthase gene ispB was disrupted, and homologues from different organisms were introduced on plasmids, generating E. coli that produced Q 7 , Q 8 , Q 9 , or Q 10 .…”

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Reproductive Fitness and Quinone Content of Caenorhabditis elegans clk-1 Mutants Fed Coenzyme Q Isoforms of Varying Length

Jonassen

Davis

Larsen

et al. 2003

Journal of Biological Chemistry

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Caenorhabditis elegans clk-1 mutants lack coenzyme Q 9 and accumulate the biosynthetic intermediate demethoxy-Q 9 . A dietary source of ubiquinone (Q) is required for larval growth and development of the gonad and germ cells. We considered that uptake of the shorter Q 8 isoform present in the Escherichia coli food may contribute to the Clk phenotypes of slowed development and reduced brood size observed when the animals are fed Q-replete E. coli. To test the effect of isoprene tail length, N2 and clk-1 animals were fed E. coli engineered to produce Q 7 , Q 8 , Q 9 , or Q 10 . Wild-type nematodes showed no change in reproductive fitness regardless of the Q n isoform fed. clk-1(e2519) fed the Q 9 diet showed increased egg production; however, this diet did not improve reproductive fitness of the clk-1(qm30) animals. Furthermore, animals with the more severe clk-1(qm30) allele become sterile and their progeny inviable when fed Q 7 -containing bacteria. The content of Q 7 in the mitochondria of clk-1 animals was decreased relative to Q 8 , suggesting less effective transport of Q 7 to the mitochondria, impaired retention, or decreased stability. Additionally, regardless of E. coli diet, clk-1(qm30) animals contain a dysfunctional dense form of mitochondria. The gonads of clk-1(qm30) worms fed Q 7 -containing food were severely shrunken and disordered. The differential fertility of clk-1 mutant nematodes fed Q isoforms may result from changes in Q localization, altered recognition by Q-binding proteins, and/or potential defects in mitochondrial function resulting from the mutant CLK-1 polypeptide itself.

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Cloning of the sdsA gene encoding solanesyl diphosphate synthase from Rhodobacter capsulatus and its functional expression in Escherichia coli and Saccharomyces cerevisiae

Cited by 62 publications

References 30 publications

A Solanesyl-diphosphate Synthase Localizes in Glycosomes of Trypanosoma cruzi

A Solanesyl-diphosphate Synthase Localizes in Glycosomes of Trypanosoma cruzi

Functional Analysis of Two Solanesyl Diphosphate Synthases fromArabidopsis thaliana

Reproductive Fitness and Quinone Content of Caenorhabditis elegans clk-1 Mutants Fed Coenzyme Q Isoforms of Varying Length

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