The glnA gene of the extremely thermophilic eubacterium Thermotoga maritima: cloning, primary structure, and expression in Escherichia coli

Sanangelantoni, Anna Maria; Forlani, Giuseppe; Ambroselli, F.; Cammarano, Piero; Tiboni, Orsola

doi:10.1099/00221287-138-2-383

Cited by 17 publications

(9 citation statements)

References 48 publications

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“…However, certain distinctive sequence signatures (Fig. 2) the GSI species examined (48,51,63), strongly support the root placement inferred from both DNA sequence-distance (Fig. 3A) and maximum-likelihood (Fig.…”

Section: Discussionsupporting

confidence: 63%

“…2), one group comprising all three archaea, Thermotoga species, and the low-G+C gram-positive bacteria, and the other encompassing S. coelicolor, cyanobacteria, and proteobacteria. Notably, these signatures also separate the GSIs whose activity is modulated by reversible adenylylation of the subunits (cyanobacteria, proteobacteria, S. coelicolor) from those whose subunits are not adenylylated (21,48,51,63).…”

Section: Asp Pgslelalealendhafltdtgvftedfiqnwidyklanevkqm-----------mentioning

confidence: 99%

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Cloning and sequencing of the gene encoding glutamine synthetase I from the archaeum Pyrococcus woesei: anomalous phylogenies inferred from analysis of archaeal and bacterial glutamine synthetase I sequences

1993

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The gene ginA encoding glutamine synthetase I (GSI) from the archaeum Pyrococcus woesei was cloned and sequenced with the Sudfolobus solfataricus ginA gene as the probe. An operon reading frame of 448 amino acids was identified within a DNA segment of 1,528 bp. The encoded protein was 49%o identical with the GSI of Methanococcus voltae and exhibited conserved regions characteristic of the GSI family. The P. woesei GSI was aligned with available homologs from other archaea (S. soifataricus, M. voitae) and with representative sequences from cyanobacteria, proteobacteria, and gram-positive bacteria. Phylogenetic trees were constructed from both the amino acid and the nucleotide sequence alignments. In accordance with the sequence similarities, archaeal and bacterial sequences did not segregate on a phylogeny. On the basis of sequence signatures, the GSI trees could be subdivided into two ensembles. One encompassed the GSI of cyanobacteria and proteobacteria, but also that of the high-G+C gram-positive bacterium Streptomyces coelicolor (all of which are regulated by the reversible adenylylation of the enzyme subunits); the other embraced the GSI of the three archaea as well as that of the low-G+C gram-positive bacteria (Clostridium acetobutilycum, Bacillus subtilis) and Thermotoga maritima (none of which are regulated by subunit adenylylation). The GSIs of the Thermotoga and the Bacilus-Clostridium lineages shared a direct common ancestor with that of P. woesei and the methanogens and were unrelated to their homologs from cyanobacteria, proteobacteria, and S. coelicolor.The possibility is presented that the GSI gene arose among the archaea and was then laterally transferred from some early methanogen to a Thermotoga-like organism. However, the relationship of the cyanobacterial-proteobacterial GSIs to the Thermotoga GSI and the GSI of low-G+C gram-positive bacteria remains unexplained.Glutamine synthetase (GS) catalyzes the ATP-dependent synthesis of glutamine from glutamate and ammonium (NH4+). The enzyme is a multimer found in at least two forms that are only distantly related to one another (15 to 19% sequence identity) (43, 56). One of these forms (GSI), composed of 12 identical subunits (443 to 474 amino acids each), occurs in bacteria and archaea. A second form (GSII), composed of eight identical subunits (332 to 378 amino acids each), occurs in eukarya but also in members of the family Rhizobiaceae (12, 23, 53) and in certain actinomycetes (4, 18, 34, 49); these last two groups harbor both a GSI and a GSII-like enzyme. A third form of the enzyme (GSIII), composed of six identical subunits (729 amino acids each), whose sequence is unrelated to both GSI and GSII, is harbored by Bacteroides fragilis (29).Because GSI is a relatively long polypeptide chain containing both semiconserved and highly conserved regions, and because it is ubiquitous in prokaryotes, GSI subunit sequences are suited, in principle, to trace the evolution of the bacterial and the archaeal lineages.A large inventory of (eu)bacterial GSI sequ...

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Section: Discussionsupporting

confidence: 63%

Section: Asp Pgslelalealendhafltdtgvftedfiqnwidyklanevkqm-----------mentioning

confidence: 99%

Cloning and sequencing of the gene encoding glutamine synthetase I from the archaeum Pyrococcus woesei: anomalous phylogenies inferred from analysis of archaeal and bacterial glutamine synthetase I sequences

1993

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“…Accordingly, these hyperthermophiles contain biomolecules that are adapted to these elevated temperatures, both in structure and function. Cloning and expression of genes from thermophiles into mesophilic hosts demonstrated that this enhanced-stability is mainly an intrinsic feature of the enzymes (Zwickl et al, 1990;Moracci et al, 1992;Sanangelantino et al, 1992). Cloning and expression of genes from thermophiles into mesophilic hosts demonstrated that this enhanced-stability is mainly an intrinsic feature of the enzymes (Zwickl et al, 1990;Moracci et al, 1992;Sanangelantino et al, 1992).…”

Section: Introductionmentioning

confidence: 99%

An Extremely Thermostable β-Glucosidase from the Hyperthermophilic ArchaeonPyrococcus Furiosus; A Comparison with Other Glycosidases

Kengen

Stams

1994

Biocatalysis

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Pyrococcus furiosus harbours several hydrolytic enzyme activities that enable growth on a variety of polymeric substrates like proteins and polysaccharides. Recently, the organism was shown to exhibit an extremely high 8-glucosidase activity, apparently involved in hydrolysis of cellobiose. The cytoplasmic enzyme was purified to homogeneity and its properties were compared with those of other glycosidases. This comparison can be summarized as follows: i) the P-glucosidase activity in cell-free extracts is at least 100-fold higher than the P. furiosus a-glucosidase activity; ii) the P-glucosidase comprises 5% of total cell protein as to only 0.3% for the a-glucosidase; iii) with respect to substrate specifity, K,, PI, and pH-optimum the P. furiosus P-glucosidase resembles other /3-glucosidases from microorganisms from lower temperature ranges; iv) compared to these P-glucosidases the P. furiosus enzyme exhibits an enhanced general stability; v) a high similarity is observed with a P-galactolglucosidase from the hyperthermophile Sulfolobus solfataricus. vi) besides intrinsic features of the enzyme, extrinsic organic compounds appear to determine thermostability of the P-glucosidase.

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“…Oligonucleotides were synthesized with a 380A DNA synthesizer (Applied Biosystems). Specific sequencing of the LDH gene in pTLDH25 was started with a degenerated oligonucleotide corresponding to the aminoterminal sequence and the codon usage of known Thermotoga genes (Bachleitner et a1.,1989;Sanangelantoni et al, 1992;Tomschy et al, 1993). The start primer was 5'AT-GAAGGGXTTTGCXAGGGAGATGGT 3' (X = G or T).…”

Section: Dna Sequencingmentioning

confidence: 99%

The l‐lactate dehydrogenase gene of the hyperthermophilic bacterium Thermotoga maritima cloned by complementation in Escherichia coli

Ostendorp

Liebl

Schurig

et al. 1993

European Journal of Biochemistry

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The gene for a l(+)‐lactate dehydrogenase from the hyperthermophilic bacterium Thermotoga maritima was cloned by complementation of an Escherichia coli pfl. ldh mutant. The gene is part of a 4.5 kb SauIIIA fragment obtained by partial digestion of the Thermotoga genome. The DNA fragment was physically mapped and the putative Shine‐Dalgarno sequence within the non‐coding region determined. The gene contains 960 bp, including the stop codon, corresponding to 319 amino acids/subunit of the homotetrameric enzyme. Part of the amino acid sequence was confirmed by Edman degradation of peptides obtained from nanomolar quantities of the purified enzyme by tryptic digestion. A comparison of the amino acid sequenc̀e with those of known prokaryotic l‐lactate dehydrogenases reveals a high similarity, especially with the enzyme from thermophilic sources, where up to 48% identity is found. The gene was expressed as an active enzyme in a heterologous host.

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The glnA gene of the extremely thermophilic eubacterium Thermotoga maritima: cloning, primary structure, and expression in Escherichia coli

Cited by 17 publications

References 48 publications

Cloning and sequencing of the gene encoding glutamine synthetase I from the archaeum Pyrococcus woesei: anomalous phylogenies inferred from analysis of archaeal and bacterial glutamine synthetase I sequences

Cloning and sequencing of the gene encoding glutamine synthetase I from the archaeum Pyrococcus woesei: anomalous phylogenies inferred from analysis of archaeal and bacterial glutamine synthetase I sequences

An Extremely Thermostable β-Glucosidase from the Hyperthermophilic ArchaeonPyrococcus Furiosus; A Comparison with Other Glycosidases

The l‐lactate dehydrogenase gene of the hyperthermophilic bacterium Thermotoga maritima cloned by complementation in Escherichia coli

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