Nucleotide Sequence of the Arginine Deiminase Gene of <i>Mycoplasma hominis</i>

Harasawa, Ryô; Koshimizu, Kaoru; Kitagawa, Masanari; Asada, Kiyozo; Kato, Ikunoshin

doi:10.1111/j.1348-0421.1992.tb02068.x

Cited by 21 publications

(11 citation statements)

References 8 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…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: 97%

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

1996

View full text Add to dashboard Cite

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...

show abstract

Section: Resultsmentioning

confidence: 97%

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

1996

View full text Add to dashboard Cite

show abstract

“…U07185), respectively. Similarities with the Mycoplasma arginini (40), Mycoplasma hominis (19), and Halobacterium salinarium (47) arginine deiminases were considerably lower (approximately 20% identity).…”

Section: Resultsmentioning

confidence: 99%

The arginine deiminase pathway in Rhizobium etli: DNA sequence analysis and functional study of the arcABC genes

et al. 1997

View full text Add to dashboard Cite

Sequence analysis upstream of the Rhizobium etli fixLJ homologous genes revealed the presence of three open reading frames homologous to the arcABC genes of Pseudomonas aeruginosa. The P. aeruginosa arcABC genes code for the enzymes of the arginine deiminase pathway: arginine deiminase, catabolic ornithine carbamoyltransferase (cOTCase), and carbamate kinase. OTCase activities were measured in free-living R. etli cells and in bacteroids isolated from bean nodules. OTCase activity in free-living cells was observed at a different pH optimum than OTCase activity in bacteroids, suggesting the presence of two enzymes with different characteristics and different expression patterns of the corresponding genes. The characteristics of the OTCase isolated from the bacteroids were studied in further detail and were shown to be similar to the properties of the cOTCase of P. aeruginosa. The enzyme has a pH optimum of 6.8 and a molecular mass of approximately 450 kDa, is characterized by a sigmoidal carbamoyl phosphate saturation curve, and exhibits a cooperativity for carbamoyl phosphate. R. etli arcA mutants, with polar effects on arcB and arcC, were constructed by insertion mutagenesis. Bean nodules induced by arcA mutants were still able to fix nitrogen but showed a significantly lower acetylene reduction activity than nodules induced by the wild type. No significant differences in nodule dry weight, plant dry weight, and number of nodules were found between the wild type and the mutants. Determination of the OTCase activity in extracts from bacteroids revealed a strong decrease in activity of this enzyme in the arcA mutant compared to the wild-type strain. Finally, we observed that expression of an R. etli arcA-gusA fusion was strongly induced under anaerobic conditions. The degradation of arginine by the arginine deiminase pathway is widely distributed among prokaryotic organisms (6). The arginine deiminase pathway consists of the following three enzymes: arginine deiminase (EC 3.5.3.6), catabolic ornithine carbamoyltransferase (cOTCase; EC 2.1.3.3), and carbamate kinase (EC 2.7.2.2). Arginine is converted into ornithine and carbamoyl phosphate, which serves to generate ATP from ADP. The degradation of 1 mol of arginine results in the formation of 1 mol of ATP and 2 mol of ammonium.cOTCase is closely related to anabolic OTCase. Anabolic OTCase is involved in arginine biosynthesis and catalyzes the reaction opposite to that catalyzed by the cOTCase, namely the formation of citrulline from ornithine and carbamoyl phosphate.The genetics of the arginine deiminase pathway have been well studied in Pseudomonas aeruginosa. In this organism, the genes encoding the enzymes of the arginine deiminase pathway are organized in an operon, the arcDABC operon. The arcA gene encodes the arginine deiminase, the arcB gene encodes the cOTCase, and the arcC gene encodes the carbamate kinase (44, 55). The fourth gene, arcD, encodes a membrane-bound protein that is necessary for the uptake of arginine and the excretion of ornithine. This tran...

show abstract

“…The predicted amino acids of the ArcA proteins of Mycoplasma hominis and Mycoplasma arginini (13,19), two species of Mycoplasma that are phylogenetically related to M. arthritidis, were aligned, and conserved amino acid regions were identified from which primers with minimal degeneracy could be designed. The arcA forward primer (5Ј-CGCTCGAGAYTAYATHACNCCNGC-3Ј) and arcA reverse primer (5Ј-GCCTCGAGCRTTNCCCATNCC-3Ј) were used to amplify a PCR product of the expected size (1.1 kb) from M. hominis strain PG21 (3) and M. arthritidis strain PG6 (23).…”

mentioning

confidence: 99%

Construction and Use of Derivatives of Transposon Tn 4001 That Function in Mycoplasma pulmonis and Mycoplasma arthritidis

2000

View full text Add to dashboard Cite

Previous attempts to introduce transposon Tn4001 into Mycoplasma pulmonis and Mycoplasma arthritidishave not been successful, possibly due to functional failure of the transposon's gentamicin resistance determinant. Tn4001C and Tn4001T were constructed, respectively, by insertion of a chloramphenicol acetyltransferase gene and the tetM tetracycline resistance determinant into Tn4001. Both Tn4001C and Tn4001T transposed in M. pulmonis, and Tn4001T transposed in M. arthritidis. The incorporation of a Tn4001T derivative that contained lacZ into either Mycoplasma species resulted in transformants with readily detectable LacZ activity. Tn4001T may be of general utility for use as a mycoplasma cloning vehicle because tetM functions in all species of Mycoplasma examined thus far.Few strategies exist to genetically manipulate mycoplasmas. There are no plasmids known to replicate in Mycoplasma species other than Mycoplasma mycoides and Mycoplasma capricolum (15, 16). The gram-positive bacterial transposon Tn916 can transpose into the genome of virtually all species of Mycoplasma for which transformation methods have been described, and Tn4001 can transpose into a number of Mycoplasma species, including Mycoplasma pneumoniae, Mycoplasma genitalium, and Mycoplasma gallisepticum (10). Transformation of these species with plasmids containing either transposon results in insertion of the transposon into the mycoplasmal genome at any of numerous sites. Tn916 is large (18 kb) and not amenable to use as a cloning vehicle. In contrast, Tn4001 (4.7 kb) is small enough to be used as a cloning vehicle to insert genes into the mycoplasmal chromosome. SmaI and BamHI cloning sites have been introduced into Tn4001 to create Tn4001mod, which has been used as a vector in M. gallisepticum and M. pneumoniae (17, 20).Tn4001 does not function in several species of Mycoplasma. Earlier reports of transposition of Tn4001 in Mycoplasma pulmonis were incorrect, and transformation of this species with plasmids containing Tn4001 has not previously been achieved (6, 18). The inability to introduce Tn4001 into M. pulmonis by transformation could result from either lack of transposase activity or failure of the transposon's gentamicin resistance determinant to serve as a selectable marker. In the present study, we inserted alternative antibiotic resistance determinants into Tn4001 to construct transposon derivatives that function in M. pulmonis and in Mycoplasma arthritidis. One of the transposon derivatives (Tn4001T) contains the tetM gene and may be used as a broad-host-range mycoplasma vector, as indicated by the successful expression of a lacZ fusion gene in both M. pulmonis and M. arthritidis.Transformation of M. pulmonis and M. arthritidis with transposons Tn4001C and Tn4001T. To determine whether Tn4001 would function in M. pulmonis if an appropriate antibiotic resistance marker was provided and to develop additional antibiotic resistance markers for this species, a chloramphenicol acetyltransferase gene (cat) was developed for use in M. pulmoni...

show abstract

Nucleotide Sequence of the Arginine Deiminase Gene of Mycoplasma hominis

Cited by 21 publications

References 8 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

The arginine deiminase pathway in Rhizobium etli: DNA sequence analysis and functional study of the arcABC genes

Construction and Use of Derivatives of Transposon Tn 4001 That Function in Mycoplasma pulmonis and Mycoplasma arthritidis

Contact Info

Product

Resources

About