Cloning and sequence analysis of the arginine deiminase gene from Mycoplasma arginini

Kondö, Keiji; Sone, Hideyuki; Yoshida, Hiroyuki; Toida, Tatsumi; Kanatani, Kazushi; Hong, Yeong‐Man; Nishino, Norikazu; Tanaka, J

doi:10.1007/bf00280371

Cited by 35 publications

(15 citation statements)

References 16 publications

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“…In the protein sequence databases, three ADIs were found. The similarities of the ADIs of P. aeruginosa (1), Mycobacterium arginini (27,32), and Mycoplasma hominis (24) to the deduced halobacterial protein are 14.1, 14.0, and 13.4%, respectively. The sequence identities of the putative halobacterial ADI and the bacterial enzymes are much lower than those that have been found for cOTCase and CK (see above).…”

Section: Resultsmentioning

confidence: 99%

Fermentative arginine degradation in Halobacterium salinarium (formerly Halobacterium halobium): genes, gene products, and transcripts of the arcRACB gene cluster

1996

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Fermentative growth via the arginine deiminase pathway is mediated by the enzymes arginine deiminase, carbamate kinase, and catabolic ornithine transcarbamylase and by a membrane-bound arginine-ornithine antiporter. Recently we reported the characterization of catabolic ornithine transcarbamylase and the corresponding gene, arcB, from Halobacterium salinarium (formerly Halobacterium halobium). Upstream of the arcB gene, three additional open reading frames with halobacterial codon usage were found. They were identified as the arcC gene coding for carbamate kinase, the arcA gene coding for arginine deiminase, and a gene, tentatively termed arcR, coding for a putative regulatory protein. The identification of the arcC and arcA genes was verified, respectively, by heterologous expression of the enzyme in Haloferax volcanii and by protein isolation and N-terminal sequence determination of three peptides. The gene order arcRACB differs from the gene order arcDABC in Pseudomonas aeruginosa, the only other organism for which sequence information is available. Transcripts from H. salinarium cultures grown fermentatively or aerobically were characterized by Northern (RNA) blot and primer extension analyses. It was determined (i) that monocistronic transcripts corresponding to the four open reading frames exist and that there are three polycistronic transcripts, (ii) that the level of induction during fermentative growth differs for the various transcripts, and (iii) that upstream of the putative transcriptional start sites for the three structural genes there are sequences with similarities to the halobacterial consensus promoter. The data indicate that expression of the arc gene cluster and its regulation differ in H. salinarium and P. aeruginosa.The arginine deiminase pathway of fermentative arginine utilization consists of the three enzymes arginine deiminase (ADI, EC 3.5.3.6), catabolic ornithine transcarbamylase (cOTCase, EC 2.1.3.3), and carbamate kinase (CK, EC 2.7.2.2) and a membrane-bound arginine-ornithine antiporter. The degradation of arginine is coupled to the equimolar generation of ATP by substrate level phosphorylation. This pathway is found in a variety of phylogenetic groups within the domain Bacteria, e.g., Pseudomonas spp., Mycoplasma spp., Bacillus spp., and lactic bacteria (for a review, see reference 8). The pathway has been most extensively studied in Pseudomonas aeruginosa (21), the only species for which the sequences of the four genes have been determined (1,2,29). The genes are organized in an operon, and a polycistronic mRNA is transcribed from a single promoter and subsequently processed into a variety of smaller transcripts (17) in an RNase E-dependent reaction (18). The transcription of the arcDABC operon is under the positive control of the ANR protein (15, 22), a member of the FNR protein family.Within the domain Archaea, arginine-dependent anaerobic growth has only been reported for some halobacterial species (25,37). The consumption of arginine was coupled to the equimolar production o...

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Section: Resultsmentioning

confidence: 99%

Fermentative arginine degradation in Halobacterium salinarium (formerly Halobacterium halobium): genes, gene products, and transcripts of the arcRACB gene cluster

1996

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“…Multiple-sequence alignments of the M. pneumoniae proteins ArcA (Mpn304, Mpn305, and Mpn560), ArcB (Mpn306), and ArcC (Mpn307) with the functional orthologous proteins from M. fermentans (10), M. penetrans (21), M. arginini (22,23), M. hominis (24), and Pseudomonas aeruginosa (25,26) revealed that key amino acids located at specific protein positions and essential for enzymatic activity were not conserved in M. pneumoniae. It was obvious for MPN_304 and MPN_305, because these genes are truncated and encode either only the N-terminal region (MPN_304) or the C-terminal region (MPN_305) of ArcA, and also for MPN_306, which encodes only a truncated ArcB of which the first 70 N-terminal amino acids are missing (5).…”

Section: Resultsmentioning

confidence: 99%

Reconstitution of an Active Arginine Deiminase Pathway in Mycoplasma pneumoniae M129

Rechnitzer¹,

Rottem

Herrmann

2013

Infect Immun

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b Some species of the genus Mycoplasma code for the arginine deiminase pathway (ADI), which enables these bacteria to produce ATP from arginine by the successive reaction of three enzymes: arginine deiminase (ArcA), ornithine carbamoyltransferase (ArcB), and carbamate kinase (ArcC). It so far appears that independently isolated strains of Mycoplasma pneumoniae encode an almost identical truncated version of the ADI pathway in which the proteins ArcA and ArcB have lost their original enzymatic activities due to the deletion of significant regions of these proteins. To study the consequences of a functional ADI pathway, M. pneumoniae M129 was successfully transformed with the cloned functional arcA, arcB, and arcC genes from Mycoplasma fermentans. Enzymatic tests showed that while the M. pneumoniae ArcAB and ArcABC transformants possess functional arginine deiminase, ornithine carbamoyltransferase, and carbamate kinase, they were unable to grow on arginine as the sole energy source. Nevertheless, infection of a lung epithelial cell line, A549, with the M. pneumoniae transformants showed that almost 100% of the infected host cells were nonviable, while most of the lung cells infected with nontransformed M. pneumoniae were viable under the same experimental conditions. T he mycoplasmas (class Mollicutes) form a large group of prokaryotic microorganisms that are divided into nine genera with over 200 species. They are distinguished from ordinary bacteria by their small size and minute genome (0.58 to 2.20 Mb) and the total lack of a cell wall (1, 2). Phylogenetically, the mycoplasmas are related to Gram-positive bacteria, from which they developed by genome reduction (3).One criterion for classifying and characterizing mycoplasma species has been their energy source. For instance, glycolytic mycoplasmas generate energy from sugars by glycolysis, while some of the nonglycolytic mycoplasmas catabolize arginine by the arginine deiminase (ADI) pathway, consisting of three enzymes: arginine deiminase (ArcA), which hydrolyzes arginine to citrulline and ammonia; ornithine carbamoyltransferase (ArcB), which converts citrulline in the presence of phosphate to ornithine and carbamoylphosphate; and carbamate kinase (ArcC), which synthesizes ATP from carbamoylphosphate and ADP. For clarity and simplicity, throughout this article these protein and gene names are interchangeable.The presence of glycolytic and/or ADI pathways in bacteria has been analyzed in the past by selective growth conditions and enzymatic assays. Barile and coworkers (4) tested 18 Mycoplasma species, of which 10, including Mycoplasma hominis, M. arthriditis, and M. fermentans, could convert arginine to ATP by the arginine deiminase pathway. Among the ADI-negative Mycoplasma species was M. pneumoniae M129, whose complete genome has been sequenced and annotated (5). Surprisingly, an operon that contains coding DNA sequences (CDSs) with significant similarities to all three enzymes of the ADI pathway and a hypothetical arginine transporter have been described fo...

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“…The similarity with P. aeruginosa (22) and M. hominis (21) was 49 and 46%, respectively. It seems that within species the sequences are well conserved, for example there is 82% similarity between the M. arginini (18) and M. hominis (21) sequences. However, when comparing sequences between species, the similarity is markedly reduced, with 54% similarity between the M. arginini (18) and P. aeruginosa (22) deiminases for example.…”

Section: Discussionmentioning

confidence: 99%

Cloning and Expression of a Prokaryotic Enzyme, Arginine Deiminase, from a Primitive Eukaryote Giardia intestinalis

Knodler¹,

Sekyere

Stewart

et al. 1998

Journal of Biological Chemistry

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Arginine deiminase (EC 3.5.3.6) catalyzes the irreversible catabolism of arginine to citrulline in the arginine dihydrolase pathway. This pathway has been regarded as restricted to prokaryotic organisms but is an important source of energy to the primitive protozoan Giardia intestinalis. In this paper we report the cloning and expression of the arginine deiminase gene from this parasite. Degenerate oligonucleotides based on amino acid sequences of tryptic peptides from the purified protein were used to amplify a portion of the arginine deiminase gene. This was then used as a probe to screen HindIII and PstI "mini" libraries to obtain two overlapping clones that contained the arginine deiminase gene. The open reading frame encoded 581 amino acids including all of the tryptic peptides that were sequenced and corresponded to a molecular mass of 67 kDa. Northern blot analysis identified a single 1.8-kilobase transcript in both trophozoites and encysting cells. Arginine deiminase was successfully expressed in Escherichia coli and purified to homogeneity. The recombinant protein was found to have characteristics comparable with those of the native enzyme.

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Cloning and sequence analysis of the arginine deiminase gene from Mycoplasma arginini

Cited by 35 publications

References 16 publications

Fermentative arginine degradation in Halobacterium salinarium (formerly Halobacterium halobium): genes, gene products, and transcripts of the arcRACB gene cluster

Fermentative arginine degradation in Halobacterium salinarium (formerly Halobacterium halobium): genes, gene products, and transcripts of the arcRACB gene cluster

Reconstitution of an Active Arginine Deiminase Pathway in Mycoplasma pneumoniae M129

Cloning and Expression of a Prokaryotic Enzyme, Arginine Deiminase, from a Primitive Eukaryote Giardia intestinalis

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