Complete sequence of the gltA gene encoding citrate synthase in Escherichia coli

Ner, Sarbjit S.; Bhayana, Vipin; Bell, Alexander W.; Giles, Ian G.; Duckworth, Harry W.; Bloxham, David P.

doi:10.1021/bi00292a001

Cited by 59 publications

(45 citation statements)

References 27 publications

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“…After 100 randomized runs, a score of 1.9 standard deviations was assigned to the alignment shown. 23 20 ogous to the E. coli enzyme (26). X-ray crystallographic AAG* 16 12 studies ( CI72 as well as in the other citrate synthases (Fig.…”

Section: Methodsmentioning

confidence: 99%

Mitochondrial and nonmitochondrial citrate synthases in Saccharomyces cerevisiae are encoded by distinct homologous genes.

Rosenkrantz¹,

Alam²,

Kim³

et al. 1986

Mol. Cell. Biol.

101

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Saccharomyces cerevisiae contains two genes, CITI and CIT2, encoding functional citrate synthase (K.-S. Kim, M. S. Rosenkrantz, and L. Guarente, Mol. Cell. Biol. 6:1936-1942). We show here that CIT2 encodes a nonmitochondrial form of citrate synthase. The DNA sequence of CIT2 presented provides a possible explanation for why the CIT2 product, unlike the CITI product, fails to be imported into mitochondria. While the products of these two genes are highly homologous, they diverge strikingly at their amino termini. The amino terminus of the CITI primary translation product extends 39 residues beyond the amino termini of Escherichia coli and porcine citrate synthases. This extension consists of a typical mitochondrial targeting motif. The amino terminus of the CIT2 primary translation product extends 20 residues beyond the amino termini of the E. coli and porcine enzymes. The CIT2-encoded extension is not homologous to that of CITI, resulting in a nonmitochondrial localization of the product. The CIT2-encoded extension, however, does bear certain similarities to mitochondrial targeting sequences. The possible role of this sequence in targeting this CIT2 product to a nonmitochondrial organelle is discussed.

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Section: Methodsmentioning

confidence: 99%

Mitochondrial and nonmitochondrial citrate synthases in Saccharomyces cerevisiae are encoded by distinct homologous genes.

Rosenkrantz¹,

Alam²,

Kim³

et al. 1986

Mol. Cell. Biol.

101

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“…The two forms of the enzyme differ in their regulatory sensitivities; the dimer is sensitive in vitro to inhibition by ATP, whereas the multimeric form is inhibited by NADH and 2-oxoglutarate. The amino acid sequences of citrate synthase either have been determined directly from the protein or have been inferred from DNA sequences for members of the domain Eukarya (including pig [9], Tetrahymena thermophila [30], Arabidopsis thaliana [50], and Saccharomyces cerevisiae [46], for members of the domain Bacteria (including Escherichia coli [28], Acinetobacter anitratum [ 141, R. prowazekii [62], Coxiella bumetii [21], Bartonella henselae [29], Pseudomonas aeruginosa [ 151, Corynebacterium glutamicum [ 161, Bacillus coagulans [43], and Mycobacterium smegmatis [12]), and for a representative of the domain Archaea, Thermoplasma acidophilum (47). Multiple alignments of these protein sequences have revealed significant levels of similarity between intradomain Eukarya sequences (46 to 92%) and between intraphylum Proteobacteria sequences (56 to 75%); however, the interdomain levels of similarity are far lower (20 to 26%) (47).…”

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confidence: 99%

Citrate Synthase Gene Comparison, a New Tool for Phylogenetic Analysis, and Its Application for the Rickettsiae

Roux

Rydkina

Eremeeva

et al. 1997

International Journal of Systematic Bacteriology

490

366

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Using PCR and an automated laser fluorescent DNA sequencer, we amplified and sequenced a 1,234-bp fragment of the citrate synthase-encoding gene ( g U ) of 28 bacteria belonging to the genus Rickettsia. Comparative sequence analysis showed that most of the spotted fever group (SFG) rickettsiae belonged to one of two subgroups. The first subgroup included Rickettsia massiliae, strain Bar 29, Rickettsia rhipicephali, "Rickettsia aeschlimanni," and Rickettsia montana, which have been isolated only from ticks. The second subgroup was larger and included the majority of the human pathogens and also rickettsiae isolated only from ticks; the members of this subgroup were strain S, Rickettsia apicae, "Rickettsia mongolotimonae," Rickettsia sibirica, Rickettsia parkeri, Rickettsia conorii, Rickettsia rickettsii, the Thai tick typhus rickettsia, the Israeli tick typhus rickettsia, the Astrakhan fever rickettsia, "Rickettsia slovaca," and Rickettsia japonica. The sequence analysis also showed that the tick-borne organisms Rickettsia helvetica and Rickettsia australis and the mite-borne organism Rickettsia akari were associated with the SFG cluster; that Rickettsia prowazekii and Rickettsia typhi, two representatives of the typhus group, clustered together; and that Rickettsia Canada, Rickettsia bellii, and the AB bacterium probably represent three new groups. We compared the phylogenetic trees inferred from citrate synthase gene sequences and from 16s ribosomal DNA (rDNA) sequences. For rickettsial phylogeny, the citrate synthase approach was more suitable, as demonstrated by significant bootstrap values for all of the nodes except those in the larger subgroup defined above. We also compared phylogenetic analysis results obtained in a comparison of the sequences of both genes for all of the representatives of the domain Bacteria for which the gZtA sequence was determined. We believe that comparison of glL4 sequences could be a complementary approach to 16s rDNA sequencing for inferring bacterial evolution, especially when unstable phylogenetic models are obtained from ribosomal sequences because of high levels of sequence similarity between the bacteria studied.Rickettsiae belong to the order Rickettsiales, whose members were described as obligately intracellular gram-negative microorganisms (55). Several species cause disease in humans or in other vertebrate and invertebrate hosts and have a worldwide distribution. Recently, on the basis of 16s rRNA sequence similarity, Rickettsia tsutsugamushi was removed from the genus Rickettsia and was renamed Orientia tsutsugarnushi (48). The genus Rickettsia is presently divided into two groups: the typhus group (TG), which includes three species (Rickettsia prowazekii, the agent of the epidemic typhus, Rickettsia typhi, which causes murine typhus, and Rickettsia Canada, which has been isolated only from ticks), and the spotted fever group (SFG). The number of recognized members of the SFG increased following the development of an improved cell culture isolation technique (the shell...

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“…Cell culture medium (minimal essential medium) was supplemented with 2% fetal bovine sera, 10 mM HEPES (N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid) buffer, nonessential amino acids, L-glutamine, and sodium bicarbonate. (24,43); the exact nucleotide sequences of both primers were derived from the nucleotide sequence of the R. prowazekii gene. It was initially posited that sequences conserved between diverse bacterial genera would most likely also be conserved within either of the genera from which the sequences were initially derived.…”

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confidence: 99%

Genotypic identification of rickettsiae and estimation of intraspecies sequence divergence for portions of two rickettsial genes

1991

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DNA sequences from specific genes, amplified by the polymerase chain reaction technique, were used as substrata for nonisotopic restriction endonuclease fragment klgth polymorphism differentiation of rickettsial species and genotypes. The products amplified using a single pai of oligonucleotide primers (derived from a rickettsial citrate synthase gene sequence) and cleaved with restriction endonucleases were used to differentiate almost all recognized species of rickettsiae. A second set of primers was used for differentiation of al recognized species of closely related spotted fever group rickettsiae. The procedure circumvents many technical obstacles previously associated with identification of rickettsial species. Multiple amplified DNA digest patterns were used to estimate the intraspecies nucleotide sequence divergence for the genes coding for rickettsial citrate synthase and a large antigen-coding gene of the spotted fever group rickettsiae. The estimted relationships deduced from these genotypic data correlate reasonably well with established rickettsial taxonomic schemes.

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Complete sequence of the gltA gene encoding citrate synthase in Escherichia coli

Cited by 59 publications

References 27 publications

Mitochondrial and nonmitochondrial citrate synthases in Saccharomyces cerevisiae are encoded by distinct homologous genes.

Mitochondrial and nonmitochondrial citrate synthases in Saccharomyces cerevisiae are encoded by distinct homologous genes.

Citrate Synthase Gene Comparison, a New Tool for Phylogenetic Analysis, and Its Application for the Rickettsiae

Genotypic identification of rickettsiae and estimation of intraspecies sequence divergence for portions of two rickettsial genes

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