Fission yeast decaprenyl diphosphate synthase consists of Dps1 and the newly characterized Dlp1 protein in a novel heterotetrameric structure

Saiki, Ryoichi; Nagata, Ai; Uchida, Naonori; Kainou, Tomohiro; Matsuda, Hideyuki; Kawamukai, Makoto

doi:10.1046/j.1432-1033.2003.03804.x

Cited by 54 publications

(85 citation statements)

References 44 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The elution position of Coq1p is similar to that of IDH, an octamer of 304 kDa. Other polyprenyl-diphosphate synthases have been postulated to form dimers and tetramers (55,56). Coq1p elutes independently of both the large and small Coq3 complexes, and no Coq1p was detected in the high molecular mass regions by BN-PAGE (data not shown).…”

Section: Discussionmentioning

confidence: 93%

Coq3 and Coq4 Define a Polypeptide Complex in Yeast Mitochondria for the Biosynthesis of Coenzyme Q

Marbois¹,

Gin²,

Faull

et al. 2005

Journal of Biological Chemistry

120

View full text Add to dashboard Cite

Coenzyme Q (Q) is a redox active lipid essential for aerobic respiration in eukaryotes. In Saccharomyces cerevisiae at least eight mitochondrial polypeptides, designated Coq1-Coq8, are required for Q biosynthesis. Here we present physical evidence for a coenzyme Qbiosynthetic polypeptide complex in isolated mitochondria. Separation of digitonin-solubilized mitochondrial extracts in one-and two-dimensional Blue Native PAGE analyses shows that Coq3 and Coq4 polypeptides comigrate as high molecular mass complexes. Similarly, gel filtration chromatography shows that Coq1p, Coq3p, Coq4p, Coq5p, and Coq6p elute in fractions higher than expected for their respective subunit molecular masses. Coq3p, Coq4p, and Coq6p coelute with an apparent molecular mass exceeding 700 kDa. Coq3 O-methyltransferase activity, a surrogate for Q biosynthesis and complex activity, also elutes at this high molecular mass. We have determined the quinone content in lipid extracts of gel filtration fractions by liquid chromatography-tandem mass spectrometry and find that demethoxy-Q 6 is enriched in fractions with Coq3p. Co-precipitation of biotinylated-Coq3 and Coq4 polypeptide from digitoninsolubilized mitochondrial extracts shows their physical association. This study identifies Coq3p and Coq4p as defining members of a Q-biosynthetic Coq polypeptide complex. Coenzyme Q (ubiquinone or Q) 1 is a redox active lipid containing a long polyprenyl tail attached to a fully substituted benzoquinone ring. The number (n) of isoprene units in the polyprenyl tail (Q n ) is distinct in different organisms; humans produce Q 10 , Caenorhabditis elegans Q 9 , Escherichia coli Q 8 , and Saccharomyces cerevisiae Q 6 . Within the inner mitochondrial membrane Q functions in respiratory electron transport, where it transfers two electrons from either complex I or complex II to complex III (1). Q is present in almost all organisms, with the quantity roughly matching the respiratory capability of the tissue from which it is isolated (2). In addition to this role, Q has been found to act as a chain-breaking antioxidant (3) and to be involved in the electron transport chains of plasma and lysosomal membranes (4, 5). Q supplementation in humans slows the functional decline in patients with Parkinson disease (6), is useful for treating patients with respiratory chain defects (7), and has promise as a treatment in other neurodegenerative diseases (8).As observed with other redox active compounds, Q has the potential to act as a source of oxidative stress. In certain species, the amount of reactive oxygen species generated by mitochondria has been related to Q content (9, 10). These described dual roles as antioxidant and prooxidant associate Q with both extended and shortened life spans. C. elegans fed E. coli diets lacking Q 8 , and C. elegans clk-1 mutants with defects in Q biosynthesis show increased life spans, and we have proposed that the decreased levels of Q are associated with a decreased level of the Q semiquinone radical (Q . ) and a decreased propensity to...

show abstract

Section: Discussionmentioning

confidence: 93%

Coq3 and Coq4 Define a Polypeptide Complex in Yeast Mitochondria for the Biosynthesis of Coenzyme Q

Marbois¹,

Gin²,

Faull

et al. 2005

Journal of Biological Chemistry

120

View full text Add to dashboard Cite

show abstract

“…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, the activity of these proteins has been experimentally demonstrated, although catalytic subunits SpDPS1 and HsDPS1 are not functional alone (62,63). Shaded areas represent amino acids identical to those in TcSPPS.…”

Section: Multialignment Of Prenyl-diphosphate Synthases (A) and Compamentioning

confidence: 99%

A Solanesyl-diphosphate Synthase Localizes in Glycosomes of Trypanosoma cruzi

Ferella

Montalvetti

Rohloff

et al. 2006

Journal of Biological Chemistry

Self Cite

View full text Add to dashboard Cite

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.

show abstract

“…Eco_ppsA, phosphoenolpyruvate synthase; Eco_aroF FBR , feedback-inhibitionresistant (FBR) 3-deoxy-D-arabino-heptulosonate 7-phosphate (DAHP) synthase; Eco_ubiC, chorismate lyase; Spo_aro2, chorismate synthase; Spo_aro7, chorismate mutase; Sce_thmgr1, truncated 3-hydroxy-3-methylglutaryl CoA (HMG-CoA) reductase 1; Sce_mvk FBR , FBR mevalonate kinase; Spo_erg8, phosphomevalonate kinase; Spo_mvd1, diphosphomevalonate decarboxylase; dps1-dlp1, decaprenyl diphosphate synthase; ppt1, PHB-decaprenyl diphosphate transferase. CoQ 10 biosynthesis 2,19,24,26) ; thus, it was a promising organism to use with this approach.…”

Section: Discussionmentioning

confidence: 99%

“…CoQ was extracted from S. pombe as described previously. 24) Briefly, crude lipid extracts were analyzed by normal phase thin layer chromatography using authentic CoQ 10 (as the standard) and benzene on a Kieselgel 60 F 254 plate. Following UV visualization, the band containing …”

Section: Methodsmentioning

confidence: 99%

Production of CoQ10 in fission yeast by expression of genes responsible for CoQ10 biosynthesis

Moriyama

Hosono

Fujii

et al. 2015

Bioscience, Biotechnology, and Biochemistry

Self Cite

View full text Add to dashboard Cite

Coenzyme Q10 (CoQ10) is essential for energy production and has become a popular supplement in recent years. In this study, CoQ10 productivity was improved in the fission yeast Schizosaccharomyces pombe. Ten CoQ biosynthetic genes were cloned and overexpressed in S. pombe. Strains expressing individual CoQ biosynthetic genes did not produce higher than a 10% increase in CoQ10 production. In addition, simultaneous expression of all ten coq genes did not result in yield improvements. Genes responsible for the biosynthesis of p-hydroxybenzoate and decaprenyl diphosphate, both of which are CoQ biosynthesis precursors, were also overexpressed. CoQ10 production was increased by overexpression of Eco_ubiC (encoding chorismate lyase), Eco_aroFFBR (encoding 3-deoxy-d-arabino-heptulosonate 7-phosphate synthase), or Sce_thmgr1 (encoding truncated HMG-CoA reductase). Furthermore, simultaneous expression of these precursor genes resulted in two fold increases in CoQ10 production.

show abstract

Fission yeast decaprenyl diphosphate synthase consists of Dps1 and the newly characterized Dlp1 protein in a novel heterotetrameric structure

Cited by 54 publications

References 44 publications

Coq3 and Coq4 Define a Polypeptide Complex in Yeast Mitochondria for the Biosynthesis of Coenzyme Q

Coq3 and Coq4 Define a Polypeptide Complex in Yeast Mitochondria for the Biosynthesis of Coenzyme Q

A Solanesyl-diphosphate Synthase Localizes in Glycosomes of Trypanosoma cruzi

Production of CoQ10 in fission yeast by expression of genes responsible for CoQ10 biosynthesis

Contact Info

Product

Resources

About