As in Saccharomyces cerevisiae, aspartate transcarbamoylase is assembled on a multifunctional protein including a dihydroorotase-like cryptic domain in Schizosaccharomyces pombe

Lollier, Marc; Jaquet, Laurence; Nedeva, Trayana; Lacroute, François; Potier, Serge; Souciet, Jean-Luc

doi:10.1007/bf00315780

Cited by 16 publications

(8 citation statements)

References 45 publications

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“…Comparison with 25 homologous genes shows that the P. abyssi pyrB and pyrI genes present the highest homology (59.4 and 58.7%, respectively) with the presumed homologous genes from the archaeon Methanococcus jannaschii (6). With eubacterial and eukaryal ATCases, the highest identities for pyrB are obtained with the corresponding enterobacterial gene from Proteus vulgaris (54.7%) (11), E. coli (54.2%) (11), Serratia marcescens (53.0%) (11), Salmonella typhimurium (52.5%) (11), and Erwinia herbicola (52.7%) (11), followed very closely by a large series of ATCase genes from eukaryotes comprising yeasts (S. cerevisiae, S. pombe) (41,47), plants (Arabidopsis thaliana, L. esculentum, Pisum sativum sp.) (48,54,79), invertebrates (Dictyostelium discoideum, Caenorhabditis elegans) (18,81), and vertebrates (Squalus acanthias, Mesocricetus auratus, C. longicaudatus) (27,42,68).…”

Section: Resultsmentioning

confidence: 99%

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Aspartate transcarbamylase from the deep-sea hyperthermophilic archaeon Pyrococcus abyssi: genetic organization, structure, and expression in Escherichia coli

et al. 1997

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The genes coding for aspartate transcarbamylase (ATCase) in the deep-sea hyperthermophilic archaeon Pyrococcus abyssi were cloned by complementation of a pyrB Escherichia coli mutant. The sequence revealed the existence of a pyrBI operon, coding for a catalytic chain and a regulatory chain, as in Enterobacteriaceae.Comparison of primary sequences of the polypeptides encoded by the pyrB and pyrI genes with those of homologous eubacterial and eukaryotic chains showed a high degree of conservation of the residues which in E. coli ATCase are involved in catalysis and allosteric regulation. The regulatory chain shows more-extensive divergence with respect to that of E. coli and other Enterobacteriaceae than the catalytic chain. Several substitutions suggest the existence in P. abyssi ATCase of additional hydrophobic interactions and ionic bonds which are probably involved in protein stabilization at high temperatures. The catalytic chain presents a secondary structure similar to that of the E. coli enzyme. Modeling of the tridimensional structure of this chain provides a folding close to that of the E. coli protein in spite of several significant differences. Conservation of numerous pairs of residues involved in the interfaces between different chains or subunits in E. coli ATCase suggests that the P. abyssi enzyme has a quaternary structure similar to that of the E. coli enzyme. P. abyssi ATCase expressed in transgenic E. coli cells exhibited reduced cooperativity for aspartate binding and sensitivity to allosteric effectors, as well as a decreased thermostability and barostability, suggesting that in P. abyssi cells this enzyme is further stabilized through its association with other cellular components.Studies of extremophilic organisms provide information on the molecular mechanisms of biological adaptation to extreme environments. In this regard, Pyrococcus abyssi is of particular interest, being a hyperthermophilic and barophilic/barotolerant archaeon. This euryarchaebacterium, isolated from a deepsea hydrothermal vent located 2,000 m deep in the North Fiji Basin, was characterized by Erauso et al. (15,16). It is a heterotrophic and strictly anaerobic microorganism and belongs to the sulfur-metabolizing group. At atmospheric pressure, it grows at 67 to 102°C, with an optimum at 96°C. Hydrostatic pressure slightly increases the growth rate of P. abyssi as well as its optimal and maximal growth temperatures (16).Aspartate transcarbamylase (ATCase) (EC 2.1.3.2) is the first enzyme of the pyrimidine biosynthetic pathway. It catalyzes the carbamylation of the amino group of aspartate by carbamylphosphate with formation of carbamylaspartate and phosphate. ATCase is extensively studied as a model for structure-function relationships in cooperativity and allosteric regulation mechanisms, as well as for the evolution of these mechanisms from prokaryotes to humans. The structure and properties of the Escherichia coli enzyme have been studied in detail (for reviews, see references 3, 10, 26, 32, 40, and 70).This dodecame...

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Section: Resultsmentioning

confidence: 99%

“…ATCases from other eubacterial and eukaryotic organisms are composed either of only catalytic chains, as in Bacillus subtilis (39) and Lycopersicum esculentum (54), or of catalytic chains associated with other enzymes of the pyrimidine pathway, as in Pseudomonas putida (67), Saccharomyces cerevisiae and Schizosaccharomyces pombe (41,69), or mammals (10,17,68).…”

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confidence: 99%

Aspartate transcarbamylase from the deep-sea hyperthermophilic archaeon Pyrococcus abyssi: genetic organization, structure, and expression in Escherichia coli

et al. 1997

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“…We speculate that the concentrations of CTP and UTP increase in the presence of HU, which blocks RNR activity. This may lead to a feedback inhibition of aspartate transcarbamoylase, the committed step in the synthesis of pyrimidines which is known to be allosterically regulated by CTP and UTP (13,23,41). Thus, HU may lead to a reduction of de novo production of uracil (Fig.…”

Section: Discussionmentioning

confidence: 99%

Cell Division Defects of Schizosaccharomyces pombe liz1 ⁻ Mutants Are Caused by Defects in Pantothenate Uptake

et al. 2004

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The liz1؉ gene of the fission yeast Schizosaccharomyces pombe was previously identified by complementation of a mutation that causes abnormal mitosis when ribonucleotide reductase is inhibited. Liz1 has similarity to transport proteins from Saccharomyces cerevisiae, but the potential substrate and its connection to the cell division cycle remain elusive. We report here that liz1 ؉ encodes a plasma membrane-localized active transport protein for the vitamin pantothenate, the precursor of coenzyme A (CoA). Liz1 is required for pantothenate uptake at low extracellular concentrations. A lack of pantothenate uptake results in three phenotypes: (i) slow growth, (ii) delayed septation, and (iii) aberrant mitosis in the presence of hydroxyurea (HU). All three phenotypes are suppressed by high extracellular concentrations of pantothenate, where pantothenate uptake occurs by passive diffusion. liz1⌬ mutants are viable because they can synthesize pantothenate from uracil as an endogenous source. The use of uracil for both pantothenate biosynthesis and deoxyribonucleotide generation provides an explanation for the aberrant mitosis in the presence of HU. HU blocks ribonucleotide reductase, and we propose that the accumulation of ribonucleotides reduces uracil biosynthesis by feedback inhibition of aspartate transcarbamoylase. Thus, the addition of HU to liz1⌬ mutants results in a shortage of pantothenate. Because liz1⌬ mutants show striking similarities to mutants with defects in fatty acid biosynthesis, we propose that the shortage of pantothenate compromises fatty acid synthesis, resulting in slow growth and mitotic defects.

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“…The S. pombe cDNA library was constructed in the vector pFL61 by M. Minet and F. Lacroute (Lollier et al ., 1995). The S. cerevisiae and S. pombe vectors have been described in Bonnefoy et al .…”

Section: Methodsmentioning

confidence: 99%

The respiratory gene OXA1 has two fission yeast orthologues which together encode a function essential for cellular viability

Bonnefoy

Kermorgant

Groudinsky

et al. 2000

Molecular Microbiology

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SummaryThe Saccharomyces cerevisiae nuclear gene OXA1, which is conserved from prokaryotes to human, was shown to be essential for cytochrome c oxidase and F1F0±ATP synthase biogenesis. We have searched for an orthologue of OXA1 in Schizosaccharomyces pombe, another yeast that is highly diverged from S. cerevisiae and which could more closely model higher eukaryotes. In particular, S. pombe exhibits a limited growth under anaerobic conditions and is petite negative, that is it does not tolerate large deletions of its mitochondrial DNA. Surprisingly, two S. pombe cDNAs able to complement an S. cerevisiae oxa1 mutation were isolated. The corresponding genes have different chromosomal locations and intron contents. They encode distinct proteins, both sharing a weak sequence identity one with the other and with Oxa1p. A phenotypic analysis of both single inactivations demonstrates that only one gene is essential for respiration in S. pombe. However, the double inactivation is lethal. This work gives new insight into the dependence of S. pombe viability upon oxa1 function, providing evidence of a connection between petite negativity, a functional respiratory chain and F1F0±ATP synthase complex in S. pombe.

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As in Saccharomyces cerevisiae, aspartate transcarbamoylase is assembled on a multifunctional protein including a dihydroorotase-like cryptic domain in Schizosaccharomyces pombe

Cited by 16 publications

References 45 publications

Aspartate transcarbamylase from the deep-sea hyperthermophilic archaeon Pyrococcus abyssi: genetic organization, structure, and expression in Escherichia coli

Aspartate transcarbamylase from the deep-sea hyperthermophilic archaeon Pyrococcus abyssi: genetic organization, structure, and expression in Escherichia coli

Cell Division Defects of Schizosaccharomyces pombe liz1 ⁻ Mutants Are Caused by Defects in Pantothenate Uptake

The respiratory gene OXA1 has two fission yeast orthologues which together encode a function essential for cellular viability

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As in Saccharomyces cerevisiae, aspartate transcarbamoylase is assembled on a multifunctional protein including a dihydroorotase-like cryptic domain in Schizosaccharomyces pombe

Cited by 16 publications

References 45 publications

Aspartate transcarbamylase from the deep-sea hyperthermophilic archaeon Pyrococcus abyssi: genetic organization, structure, and expression in Escherichia coli

Aspartate transcarbamylase from the deep-sea hyperthermophilic archaeon Pyrococcus abyssi: genetic organization, structure, and expression in Escherichia coli

Cell Division Defects of Schizosaccharomyces pombe liz1 − Mutants Are Caused by Defects in Pantothenate Uptake

The respiratory gene OXA1 has two fission yeast orthologues which together encode a function essential for cellular viability

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Cell Division Defects of Schizosaccharomyces pombe liz1 ⁻ Mutants Are Caused by Defects in Pantothenate Uptake