Thermostability and Thermoactivity of Citrate Synthases from the Thermophilic and Hyperthermophilic Archaea, Thermoplasma acidophilum and Pyrococcus furiosus

2000 ◽  
Vol 304 (4) ◽  
pp. 657-668 ◽  
Author(s):  
Michael A. Arnott ◽  
Rebecca A. Michael ◽  
Carl R. Thompson ◽  
David W. Hough ◽  
Michael J. Danson
Keyword(s):  
Pyrococcus Furiosus ◽  
Hyperthermophilic Archaea ◽  
Thermoplasma Acidophilum

scholarly journals Genome Evolution at the Genus Level: Comparison of Three Complete Genomes of Hyperthermophilic Archaea

2001 ◽  
Vol 11 (6) ◽  
pp. 981-993
Author(s):  
Odile Lecompte ◽  
Raymond Ripp ◽  
Valérie Puzos-Barbe ◽  
Simone Duprat ◽  
Roland Heilig ◽  
...  
Keyword(s):  
Pyrococcus Furiosus ◽  
Hyperthermophilic Archaea ◽  
Trna Genes ◽  
Distance Ratio ◽  
Genomic Plasticity ◽  
Relative Contribution ◽  
Complete Genomes ◽  
Replication Terminus ◽  
Living Organisms ◽  
High Level

We have compared three complete genomes of closely related hyperthermophilic species of Archaea belonging to thePyrococcus genus: Pyrococcus abyssi, Pyrococcus horikoshii, and Pyrococcus furiosus. At the genomic level, the comparison reveals a differential conservation among four regions of the Pyrococcus chromosomes correlated with the location of genetic elements mediating DNA reorganization. This discloses the relative contribution of the major mechanisms that promote genomic plasticity in these Archaea, namely rearrangements linked to the replication terminus, insertion sequence-mediated recombinations, and DNA integration within tRNA genes. The combination of these mechanisms leads to a high level of genomic plasticity in these hyperthermophilic Archaea, at least comparable to the plasticity observed between closely related bacteria. At the proteomic level, the comparison of the threePyrococcus species sheds light on specific selection pressures acting both on their coding capacities and evolutionary rates. Indeed, thanks to two independent methods, the “reciprocal best hits“ approach and a new distance ratio analysis, we detect the false orthology relationships within the Pyrococcus lineage. This reveals a high amount of differential gains and losses of genes since the divergence of the three closely related species. The resulting polymorphism is probably linked to an adaptation of these free-living organisms to differential environmental constraints. As a corollary, we delineate the set of orthologous genes shared by the three species, that is, the genes that may characterize the Pyrococcus genus. In this conserved core, the amino acid substitution rate is equal between P. abyssi and P. horikoshii for most of their shared proteins, even for fast-evolving ones. In contrast, strong discrepancies exist among the substitution rates observed in P. furiosus relative to the two other species, which is in disagreement with the molecular clock hypothesis.

scholarly journals Novel Type of ADP-Forming Acetyl Coenzyme A Synthetase in Hyperthermophilic Archaea: Heterologous Expression and Characterization of Isoenzymes from the Sulfate Reducer Archaeoglobus fulgidus and the Methanogen Methanococcus jannaschii

2002 ◽  
Vol 184 (3) ◽  
pp. 636-644 ◽  
Author(s):  
Meike Musfeldt ◽  
Peter Schönheit
Keyword(s):  
Coenzyme A ◽  
Pyrococcus Furiosus ◽  
Sequence Similarity ◽  
Hyperthermophilic Archaea ◽  
Archaeoglobus Fulgidus ◽  
Methanococcus Jannaschii ◽  
Acetyl Coa ◽  
Acetyl Coenzyme A ◽  
Acetyl Coenzyme ◽  
Α And Β Subunits

ABSTRACT Acetyl coenzyme A (CoA) synthetase (ADP forming) (ACD) represents a novel enzyme of acetate formation and energy conservation (acetyl-CoA + ADP + Pi ⇌ acetate + ATP + CoA) in Archaea and eukaryotic protists. The only characterized ACD in archaea, two isoenzymes from the hyperthermophile Pyrococcus furiosus, constitute 145-kDa heterotetramers (α2, β2). The coding genes for the α and β subunits are located at different sites in the P. furiosus chromosome. Based on significant sequence similarity of the P. furiosus genes, five open reading frames (ORFs) encoding putative ACD were identified in the genome of the hyperthermophilic sulfate-reducing archaeon Archaeoglobus fulgidus and one ORF was identified in the hyperthermophilic methanogen Methanococcus jannaschii. The ORFs constitute fusions of the homologous P. furiosus genes encoding the α and β subunits. Two ORFs, AF1211 and AF1938, of A. fulgidus and ORF MJ0590 of M. jannaschii were cloned and functionally overexpressed in Escherichia coli. The purified recombinant proteins were characterized as distinctive isoenzymes of ACD with different substrate specificities. In contrast to the Pyrococcus ACD, the ACDs of Archaeoglobus and Methanococcus constitute homodimers of about 140 kDa composed of two identical 70-kDa subunits, which represent fusions of the homologous P. furiosus α and β subunits in an αβ (AF1211 and MJ0590) or βα (AF1938) orientation. The data indicate that A. fulgidus and M. jannaschii contains a novel type of ADP-forming acetyl-CoA synthetase in Archaea, in which the subunit polypeptides and their coding genes are fused.

scholarly journals A First Analysis of Metallome Biosignatures of Hyperthermophilic Archaea

Archaea ◽  
2012 ◽  
Vol 2012 ◽  
pp. 1-12 ◽  
Author(s):  
Vyllinniskii Cameron ◽  
Christopher H. House ◽  
Susan L. Brantley
Keyword(s):  
Trace Metal ◽  
Pyrococcus Furiosus ◽  
Hyperthermophilic Archaea ◽  
Metal Concentrations ◽  
E Coli ◽  
Reverse Pattern ◽  
Icp Ms ◽  
Metal Contents ◽  
Mesophilic Bacterium ◽  
Aerobic And Anaerobic

To date, no experimental data has been reported for the metallome of hyperthermophilic microorganisms although their metal requirements for growth are known to be unique. Here, experiments were conducted to determine (i) cellular trace metal concentrations of the hyperthermophilic ArchaeaMethanococcus jannaschiiandPyrococcus furiosus, and (ii) a first estimate of the metallome for these hyperthermophilic species via ICP-MS. The metal contents of these cells were compared to parallel experiments using the mesophilic bacteriumEscherichia coligrown under aerobic and anaerobic conditions. Fe and Zn were typically the most abundant metals in cells. Metal concentrations forE. coligrown aerobically decreased in the order Fe > Zn > Cu > Mo > Ni > W > Co. In contrast,M. jannaschiiandP. furiosusshow almost the reverse pattern with elevated Ni, Co, and W concentrations. Of the three organisms, a biosignature is potentially demonstrated for the methanogenM. jannaschiithat may, in part, be related to the metallome requirements of methanogenesis. The bioavailability of trace metals more than likely has varied through time. If hyperthermophiles are very ancient, then the trace metal patterns observed here may begin to provide some insights regarding Earth's earliest cells and in turn, early Earth chemistry.

Metabolic channelling of carbamoyl phosphate in the hyperthermophilic archaeon Pyrococcus furiosus: dynamic enzyme–enzyme interactions involved in the formation of the channelling complex

2004 ◽  
Vol 32 (2) ◽  
pp. 306-309 ◽  
Author(s):  
J. Massant ◽  
N. Glansdorff
Keyword(s):  
Protein Interactions ◽  
Pyrococcus Furiosus ◽  
Hyperthermophilic Archaea ◽  
Physical Interaction ◽  
Protein Protein Interactions ◽  
Carbamoyl Phosphate ◽  
Molecular Physiology ◽  
Metabolic Channelling ◽  
Aspartate Carbamoyltransferase ◽  
Carbamate Kinase

Protection of thermolabile metabolites and coenzymes is a somewhat neglected but essential aspect of the molecular physiology of hyperthermophiles. Detailed information about the mechanisms used by thermophiles to protect these thermolabile metabolites and coenzymes is still scarce. A case in point is CP (carbamoyl phosphate), a precursor of pyrimidines and arginine, which is an extremely labile and potentially toxic intermediate. Recently we obtained the first evidence for a physical interaction between two hyperthermophilic enzymes for which kinetic evidence had suggested that these enzymes channel a highly thermolabile and potentially toxic intermediate. By physically interacting with each other, CKase (carbamate kinase) and OTCase (ornithine carbamoyltransferase) prevent thermodenaturation of CP in the aqueous cytoplasmic environment. The CP channelling complex involving CKase and OTCase or ATCase (aspartate carbamoyltransferase), identified in hyperthermophilic archaea, provides a good model system to investigate the mechanism of metabolic channelling and the molecular basis of protein–protein interactions in the physiology of extreme thermophiles.

scholarly journals Positive, Neutral and Negative Interactions in Cocultures between Pyrococcus furiosus and Different Methanogenic Archaea

2012 ◽  
Vol 5 ◽  
pp. MBI.S8516 ◽  
Author(s):  
Agnes Weiner ◽  
Simone Schopf ◽  
Gerhard Wanner ◽  
Alexander Probst ◽  
Reinhard Wirth
Keyword(s):  
Pyrococcus Furiosus ◽  
Model Organism ◽  
Methanogenic Archaea ◽  
Hyperthermophilic Archaea ◽  
Positive Interactions ◽  
Metabolic Intermediates ◽  
Positive Effects ◽  
Methanopyrus Kandleri ◽  
Inhibitory Interactions ◽  
Microscopic Techniques

The model organism Pyrococcus furiosus has recently been reported to interact with Methanopyrus kandleri in coculture, suggesting a H2 symbiosis. In the current study we further investigated this hypothesis by growing P. furiosus with four other hyperthermophilic methanogens providing evidence that the organisms did not only exert positive effects ( P. furiosus/ Methanocaldococcus villosus and P. furiosus/ Methanocaldococcus infernus) on each other, but also neutral ( P. furiosus/ Methanocaldococcus jannaschii) and even inhibitory interactions ( P. furiosus/ Methanotorris igneus) were detected suggesting interspecies relationships not only based on H2 symbiosis. Using various microscopic techniques we further analyzed the coculture with the highest positive interactions ( P. furiosus/ M. villosus) concerning its growth behavior on various surfaces, which turned out to be in stark contrast to the previous reported coculture of P. furiosus/ M. kandleri. This communication provides new insights into possible interactions of extremophilic Archaea in cocultures and again raises the question if and how hyperthermophilic Archaea communicate besides metabolic intermediates like H2.

scholarly journals Novel Intact Polar and Core Lipid Compositions in the Pyrococcus Model Species, P. furiosus and P. yayanosii, Reveal the Largest Lipid Diversity Amongst Thermococcales

Biomolecules ◽  
2020 ◽  
Vol 10 (6) ◽  
pp. 830
Author(s):  
Maxime Tourte ◽  
Vanessa Kuentz ◽  
Philippe Schaeffer ◽  
Vincent Grossi ◽  
Anais Cario ◽  
...  
Keyword(s):  
Lipid Composition ◽  
Pyrococcus Furiosus ◽  
Phosphatidyl Inositol ◽  
Structural Diversity ◽  
Hyperthermophilic Archaea ◽  
Membrane Composition ◽  
Acid Methanolysis ◽  
First Time ◽  
Pyrococcus Yayanosii

Elucidating the lipidome of Archaea is essential to understand their tolerance to extreme environmental conditions. Previous characterizations of the lipid composition of Pyrococcus species, a model genus of hyperthermophilic archaea belonging to the Thermococcales order, led to conflicting results, which hindered the comprehension of their membrane structure and the putative adaptive role of their lipids. In an effort to clarify the lipid composition data of the Pyrococcus genus, we thoroughly investigated the distribution of both the core lipids (CL) and intact polar lipids (IPL) of the model Pyrococcus furiosus and, for the first time, of Pyrococcus yayanosii, the sole obligate piezophilic hyperthermophilic archaeon known to date. We showed a low diversity of IPL in the lipid extract of P. furiosus, which nonetheless allowed the first report of phosphatidyl inositol-based glycerol mono- and trialkyl glycerol tetraethers. With up to 13 different CL structures identified, the acid methanolysis of Pyrococcus furiosus revealed an unprecedented CL diversity and showed strong discrepancies with the IPL compositions reported here and in previous studies. By contrast, P. yayanosii displayed fewer CL structures but a much wider variety of polar heads. Our results showed severe inconsistencies between IPL and CL relative abundances. Such differences highlight the diversity and complexity of the Pyrococcus plasma membrane composition and demonstrate that a large part of its lipids remains uncharacterized. Reassessing the lipid composition of model archaea should lead to a better understanding of the structural diversity of their lipidome and of their physiological and adaptive functions.

scholarly journals Characterization of Ten Heterotetrameric NDP-Dependent Acyl-CoA Synthetases of the Hyperthermophilic ArchaeonPyrococcus furiosus

Archaea ◽  
2014 ◽  
Vol 2014 ◽  
pp. 1-9 ◽  
Author(s):  
Joseph W. Scott ◽  
Farris L. Poole ◽  
Michael W. W. Adams
Keyword(s):  
Amino Acids ◽  
Phosphoenolpyruvate Carboxykinase ◽  
Pyrococcus Furiosus ◽  
Hyperthermophilic Archaea ◽  
E Coli ◽  
Proteomic Data ◽  
Genes Encoding ◽  
Derivatives Of ◽  
Dependent Processes

The hyperthermophilic archaeonPyrococcus furiosusgrows by fermenting peptides and carbohydrates to organic acids. In the terminal step, acyl-CoA synthetase (ACS) isoenzymes convert acyl-CoA derivatives to the corresponding acid and conserve energy in the form of ATP. ACS1 and ACS2 were previously purified fromP. furiosusand haveα2β2structures but the genome contains genes encoding three additionalα-subunits. The ten possible combinations ofαandβgenes were expressed inE. coliand each resulted in stable and activeα2β2isoenzymes. Theα-subunit of each isoenzyme determined CoA-based substrate specificity and between them they accounted for the CoA derivatives of fourteen amino acids. Theβ-subunit determined preference for adenine or guanine nucleotides. The GTP-generating isoenzymes are proposed to play a role in gluconeogenesis by producing GTP for GTP-dependent phosphoenolpyruvate carboxykinase and for other GTP-dependent processes. Transcriptional and proteomic data showed that all ten isoenzymes are constitutively expressed indicating that both ATP and GTP are generated from the metabolism of most of the amino acids. A phylogenetic analysis showed that the ACSs ofP. furiosusand other members of the Thermococcales are evolutionarily distinct from those found throughout the rest of biology, including those of other hyperthermophilic archaea.

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