Occurrence of several genes encoding putative reductive dehalogenases inDesulfitobacterium hafniense/frappieriandDehalococcoides ethenogenes

2002 ◽  
Vol 48 (8) ◽  
pp. 697-706 ◽  
Author(s):  
Richard Villemur ◽  
Maude Saucier ◽  
Annie Gauthier ◽  
Réjean Beaudet
Keyword(s):  
Open Reading Frames ◽  
Reductive Dehalogenation ◽  
Cosmid Library ◽  
Gene Arrangement ◽  
Gene Encoding ◽  
Conserved Sequence ◽  
Dehalococcoides Ethenogenes ◽  
Degenerate Oligonucleotide ◽  
Genes Encoding ◽  
Desulfitobacterium Hafniense

Desulfitobacterium frappieri PCP-1 has the capacity to dehalogenate several halogenated aromatic compounds by reductive dehalogenation, however, the genes encoding the enzymes involved in such processes have not yet been identified. Using a degenerate oligonucleotide corresponding to a conserved sequence of CprA/PceA reductive dehalogenases, a cprA-like gene fragment was amplified by PCR from this bacterial strain. A Delfitobacterium frappieri PCP-1 cosmid library was screened with the PCR product, allowing the cloning and sequencing of a 1.9-kb fragment. This fragment contains a nucleic acid sequence identical to one genomic contig of Desulfitobacterium hafniense, a bacterium closely related to Delfitobacterium frappieri that is also involved in reductive dehalogenation. Other genes related to the Desulfitobacterium dehalogenans cpr locus were identified in this contig. Interestingly, the gene arrangement shows the presence of two copies of cprA-, cprB-, cprC-, cprD-, cprK-, and cprT-related genes, suggesting that gene duplication occurred within this chromosomic region. The screening of Delfitobacterium hafniense genomic contigs with a CprA-deduced amino acid sequence revealed two other cprA-like genes. Microbial genomes available in gene databases were also analyzed for sequences related to CprA/PceA. Two open reading frames encoding other putative reductive dehalogenases in Delfitobacterium hafniense contigs were detected, along with 17 in the Dehalococcoides ethenogenes genome, a bacterium involved in the reductive dehalogenation of tetrachloroethene to ethene. The fact that several gene encoding putative reductive dehalogenases exist in Delfitobacterium hafniense, probably in other members of the genus Desulfitobacterium, and in Dehalococcoides ethenogenes suggests that these bacteria use distinct but related enzymes to achieve the dehalogenation of several chlorinated compounds.Key words: Desulfitobacterium, reductive dehalogenases, halorespiration, chlorinated compounds, gene family.

scholarly journals Replication Region Analysis Reveals Non-lambdoid Shiga Toxin Converting Bacteriophages

2021 ◽  
Vol 12 ◽  
Author(s):  
Ann-Katrin Llarena ◽  
Marina Aspholm ◽  
Kristin O’Sullivan ◽  
Grzegorz Wêgrzyn ◽  
Toril Lindbäck
Keyword(s):  
Shiga Toxin ◽  
Gene Arrangement ◽  
Gene Encoding ◽  
Lambdoid Phage ◽  
Virulence Potential ◽  
Genes Encoding ◽  
Large Outbreak ◽  
Replication Region ◽  
Family Based ◽  
Novel Strategy

Shiga toxin is the major virulence factor of enterohemorrhagic Escherichia coli (EHEC), and the gene encoding it is carried within the genome of Shiga toxin-converting phages (Stx phages). Numerous Stx phages have been sequenced to gain a better understanding of their contribution to the virulence potential of EHEC. The Stx phages are classified into the lambdoid phage family based on similarities in lifestyle, gene arrangement, and nucleotide sequence to the lambda phages. This study explores the replication regions of non-lambdoid Stx phages that completely lack the O and P genes encoding the proteins involved in initiating replication in the lambdoid phage genome. Instead, they carry sequences encoding replication proteins that have not been described earlier, here referred to as eru genes (after EHEC phage replication unit genes). This study identified three different types of Eru-phages, where the Eru1-type is carried by the highly pathogenic EHEC strains that caused the Norwegian O103:H25 outbreak in 2006 and the O104:H4 strain that caused the large outbreak in Europe in 2011. We show that Eru1-phages exhibit a less stable lysogenic state than the classical lambdoid Stx phages. As production of phage particles is accompanied by production of Stx toxin, the Eru1-phage could be associated with a high-virulence phenotype of the host EHEC strain. This finding emphasizes the importance of classifying Stx phages according to their replication regions in addition to their Stx-type and could be used to develop a novel strategy to identify highly virulent EHEC strains for improved risk assessment and management.

scholarly journals Ferrous Iron- and Sulfur-Induced Genes in Sulfolobus metallicus

2007 ◽  
Vol 73 (8) ◽  
pp. 2491-2497 ◽  
Author(s):  
Stephan Bathe ◽  
Paul R. Norris
Keyword(s):  
Reverse Transcription ◽  
Ferrous Iron ◽  
Subtractive Hybridization ◽  
Open Reading Frames ◽  
Gene Encoding ◽  
Reverse Transcription Pcr ◽  
Genes Encoding ◽  
Induced Genes ◽  
Reading Frames ◽  
Sulfur Oxygenase Reductase

ABSTRACT Genes of Sulfolobus metallicus that appeared to be upregulated in relation to growth on either ferrous iron or sulfur were identified using subtractive hybridization of cDNAs. The genes upregulated during growth on ferrous iron were found in a cluster, and most were predicted to encode membrane proteins. Quantitative reverse transcription-PCR of cDNA showed upregulation of most of these genes during growth on ferrous iron and pyrite compared to results during growth on sulfur. The highest expression levels observed included those for genes encoding proteins with similarities to cytochrome c oxidase subunits and a CbsA-like cytochrome. The genes identified here that may be involved in oxidation of ferrous iron by S. metallicus are termed fox genes. Of three available genomes of Sulfolobus species (S. tokodaii, S. acidocaldarius, and S. solfataricus), only that of S. tokodaii has a cluster of highly similar open reading frames, and only S. tokodaii of these three species was also able to oxidize ferrous iron. A gene encoding sulfur oxygenase-reductase was identified as the source of the dominant transcript in sulfur-grown cells of S. metallicus, with the predicted protein showing high identities to the previously described examples from S. tokodaii and species of Acidianus.

scholarly journals Cytochrome P460 Genes from the MethanotrophMethylococcus capsulatus Bath

1998 ◽  
Vol 180 (24) ◽  
pp. 6440-6445 ◽  
Author(s):  
David J. Bergmann ◽  
James A. Zahn ◽  
Alan B. Hooper ◽  
Alan A. DiSpirito
Keyword(s):  
Sequence Similarity ◽  
Immunoblot Analysis ◽  
Open Reading Frames ◽  
Binding Motif ◽  
Type I ◽  
Methylococcus Capsulatus ◽  
Gene Encoding ◽  
Degenerate Oligonucleotide ◽  
Apparent Similarity ◽  
Ammonia Oxidizing Bacterium

ABSTRACT P460 cytochromes catalyze the oxidation of hydroxylamine to nitrite. They have been isolated from the ammonia-oxidizing bacteriumNitrosomonas europaea (R. H. Erickson and A. B. Hooper, Biochim. Biophys. Acta 275:231–244, 1972) and the methane-oxidizing bacterium Methylococcus capsulatus Bath (J. A. Zahn et al., J. Bacteriol. 176:5879–5887, 1994). A degenerate oligonucleotide probe was synthesized based on the N-terminal amino acid sequence of cytochrome P460 and used to identify a DNA fragment from M. capsulatus Bath that containscyp, the gene encoding cytochrome P460. cyp is part of a gene cluster that contains three open reading frames (ORFs), the first predicted to encode a 59,000-Da membrane-bound polypeptide, the second predicted to encode a 12,000-Da periplasmic protein, and the third (cyp) encoding cytochrome P460. The products of the first two ORFs have no apparent similarity to any proteins in the GenBank database. The overall sequence similarity of the P460 cytochromes from M. capsulatus Bath and N. europaea was low (24.3% of residues identical), although short regions of conserved residues are present in the two proteins. Both cytochromes have a C-terminal, c-heme binding motif (CXXCH) and a conserved lysine residue (K61) that may provide an additional covalent cross-link to the heme (D. M. Arciero and A. B. Hooper, FEBS Lett. 410:457–460, 1997). Gene probing usingcyp indicated that a cytochrome P460 similar to that fromM. capsulatus Bath may be present in the type II methanotrophs Methylosinus trichosporium OB3b andMethylocystis parvus OBBP but not in the type I methanotrophs Methylobacter marinus A45,Methylomicrobium albus BG8, and Methylomonassp. strains MN and MM2. Immunoblot analysis with antibodies against cytochrome P460 from M. capsulatus Bath indicated that the expression level of cytochrome P460 was not affected either by expression of the two different methane monooxygenases or by addition of ammonia to the culture medium.

scholarly journals Physiological Control and Regulation of the Rhodobacter capsulatus cbb Operons

1998 ◽  
Vol 180 (16) ◽  
pp. 4258-4269 ◽  
Author(s):  
George C. Paoli ◽  
Padungsri Vichivanives ◽  
F. Robert Tabita
Keyword(s):  
Rhodobacter Capsulatus ◽  
Pentose Phosphate ◽  
Open Reading Frames ◽  
Physiological Control ◽  
Gene Encoding ◽  
Bisphosphate Carboxylase ◽  
Promoter Fusion ◽  
Fructose 1,6 Bisphosphatase ◽  
Mutant Strains ◽  
Genes Encoding

ABSTRACT The genes encoding enzymes of the Calvin-Benson-Bassham (CBB) reductive pentose phosphate pathway in Rhodobacter capsulatus are organized in at least two operons, each preceded by a separate cbbR gene, encoding potential LysR-type transcriptional activators. As a prelude to studies ofcbb gene regulation in R. capsulatus, the nucleotide sequence of a 4,537-bp region, which includedcbbR II, was determined. This region contained the following open reading frames: a partial pgmgene (encoding phosphoglucomutase) and a complete qorgene (encoding NADPH:quinone oxidoreductase), followed by cbbR II, cbbF (encoding fructose 1,6-bisphosphatase), cbbP (encoding phosphoribulokinase), and part of cbbT (encoding transketolase). Physiological control of the CBB pathway and regulation of the R. capsulatus cbb genes were studied by using a combination of mutant strains and promoter fusion constructs. Characterization of mutant strains revealed that either form I or form II ribulose 1,5-bisphosphate carboxylase/oxygenase (RubisCO), encoded by the cbbLS andcbbM genes, respectively, could support photoheterotrophic and autotrophic growth. A strain with disruptions in both cbbL and cbbM could not grow autotrophically and grew photoheterotrophically only when dimethyl sulfoxide was added to the culture medium. Disruption ofcbbP resulted in a strain that did not synthesize form II RubisCO and had a phenotype similar to that observed in the RubisCO-minus strain, suggesting that there is only onecbbP gene in R. capsulatus and that this gene is cotranscribed with cbbM. Analysis of RubisCO activity and synthesis in strains with disruptions in eithercbbR I orcbbR II, and β-galactosidase determinations from wild-type and mutant strains containing cbb Ip- andcbb IIp-lacZ fusion constructs, indicated that the cbb I andcbb II operons of R. capsulatus are within separate CbbR regulons.

scholarly journals Colocation of Genes Encoding a tRNA-mRNA Hybrid and a Putative Signaling Peptide on Complementary Strands in the Genome of the Hyperthermophilic Bacterium Thermotoga maritima

2006 ◽  
Vol 188 (19) ◽  
pp. 6802-6807 ◽  
Author(s):  
Clemente I. Montero ◽  
Derrick L. Lewis ◽  
Matthew R. Johnson ◽  
Shannon B. Conners ◽  
Elizabeth A. Nance ◽  
...  
Keyword(s):  
Thermotoga Maritima ◽  
Normal Growth ◽  
Growth Conditions ◽  
Open Reading Frames ◽  
Transcriptional Responses ◽  
Gene Encoding ◽  
Hyperthermophilic Bacterium ◽  
Genes Encoding ◽  
Original Genome ◽  
Opposite Strand

ABSTRACT In the genome of the hyperthermophilic bacterium Thermotoga maritima, TM0504 encodes a putative signaling peptide implicated in population density-dependent exopolysaccharide formation. Although not noted in the original genome annotation, TM0504 was found to colocate, on the opposite strand, with the gene encoding ssrA, a hybrid of tRNA and mRNA (tmRNA), which is involved in a trans-translation process related to ribosome rescue and is ubiquitous in bacteria. Specific DNA probes were designed and used in real-time PCR assays to follow the separate transcriptional responses of the colocated open reading frames (ORFs) during transition from exponential to stationary phase, chloramphenicol challenge, and syntrophic coculture with Methanococcus jannaschii. TM0504 transcription did not vary under normal growth conditions. Transcription of the tmRNA gene, however, was significantly up-regulated during chloramphenicol challenge and in T. maritima bound in exopolysaccharide aggregates during methanogenic coculture. The significance of the colocation of ORFs encoding a putative signaling peptide and tmRNA in T. maritima is intriguing, since this overlapping arrangement (tmRNA associated with putative small ORFs) was found to be conserved in at least 181 bacterial genomes sequenced to date. Whether peptides related to TM0504 in other bacteria play a role in quorum sensing is not yet known, but their ubiquitous colocalization with respect to tmRNA merits further examination.

scholarly journals Identification of a Novel Dioxygenase Involved in Metabolism of o-Xylene, Toluene, and Ethylbenzene by Rhodococcus sp. Strain DK17

2004 ◽  
Vol 70 (12) ◽  
pp. 7086-7092 ◽  
Author(s):  
Dockyu Kim ◽  
Jong-Chan Chae ◽  
Gerben J. Zylstra ◽  
Young-Soo Kim ◽  
Seong-Ki Kim ◽  
...  
Keyword(s):  
Aromatic Ring ◽  
Single Point ◽  
Open Reading Frames ◽  
Gene Encoding ◽  
Catabolic Genes ◽  
Genes Encoding ◽  
Sulfur Protein ◽  
Benzene Toluene ◽  
Iron Sulfur Protein ◽  
Dioxygenase Genes

ABSTRACT Rhodococcus sp. strain DK17 is able to grow on o-xylene, benzene, toluene, and ethylbenzene. DK17 harbors at least two megaplasmids, and the genes encoding the initial steps in alkylbenzene metabolism are present on the 330-kb pDK2. The genes encoding alkylbenzene degradation were cloned in a cosmid clone and sequenced completely to reveal 35 open reading frames (ORFs). Among the ORFs, we identified two nearly exact copies (one base difference) of genes encoding large and small subunits of an iron sulfur protein terminal oxygenase that are 6 kb apart from each other. Immediately downstream of one copy of the dioxygenase genes (akbA1a and akbA2a ) is a gene encoding a dioxygenase ferredoxin component (akbA3), and downstream of the other copy (akbA1b and akbA2b ) are genes putatively encoding a meta-cleavage pathway. RT-PCR experiments show that the two copies of the dioxygenase genes are operonic with the downstream putative catabolic genes and that both operons are induced by o-xylene. When expressed in Escherichia coli, AkbA1a-AkbA2a-AkbA3 transformed o-xylene into 2,3- and 3,4-dimethylphenol. These were apparently derived from an unstable o-xylene cis-3,4-dihydrodiol, which readily dehydrates. This indicates a single point of attack of the dioxygenase on the aromatic ring. In contrast, attack of AkbA1a-AkbA2a-AkbA3 on ethylbenzene resulted in the formation of two different cis-dihydrodiols resulting from an oxidation at the 2,3 and the 3,4 positions on the aromatic ring, respectively.

scholarly journals The tet39 Determinant and the msrE-mphE Genes in Acinetobacter Plasmids Are Each Part of Discrete Modules Flanked by Inversely Oriented pdif (XerC-XerD) Sites

2017 ◽  
Vol 61 (8) ◽  
Author(s):  
Grace A. Blackwell ◽  
Ruth M. Hall
Keyword(s):  
Insertion Sequence ◽  
Insertion Site ◽  
Tetracycline Resistance ◽  
Open Reading Frames ◽  
Gene Encoding ◽  
Content Type ◽  
System A ◽  
Genes Encoding ◽  
Putative Toxin ◽  
Reading Frames

ABSTRACT The tet39 tetracycline resistance determinant and the macrolide resistance genes msrE and mphE were found in an 18.2-kb plasmid, pS30-1, recovered from a global clone 2 (GC2) Acinetobacter baumannii isolate from Singapore, that conferred resistance to tetracycline and erythromycin. pS30-1 also contains mobA and mobC genes encoding MOBQ family proteins, but attempts to mobilize pS30-1 utilizing a coresident conjugative repAci6 plasmid were unsuccessful. Eight pdif sites, consisting of inversely oriented binding sites for the XerC and XerD recombinases separated by 6 bp, were detected in pS30-1. The tet39 determinant and the msrE-mphE gene pair are each surrounded by two pdif sites in inverse orientation. Identical regions in different contexts and many previously unnoticed pdif sites were found in a number of different plasmids in GenBank, showing that the tet39 and msrE-mphE dif modules are mobile. A putative toxin/antitoxin system, a gene encoding a serine recombinase, and open reading frames of unknown function were also part of dif modules in pS30-1. In general, modules with internal XerC or XerD sites alternate. Two copies of ISAjo2-1 (94% identical to ISAjo2) in pS30-1 were inserted 5 bp from a XerC site, and this appears to be the preferred insertion site for this insertion sequence (IS) group. Apparently, Acinetobacter plasmids exploit the Acinetobacter XerC-XerD recombinases to mobilize DNA units containing resistance and other genes, via an uncharacterized mechanism. The tet39 and msrE-mphE dif modules add to the oxa24 module and the oxa58 module redefined here, bringing the total of resistance gene-containing dif modules in Acinetobacter plasmids to four.

scholarly journals Chlamydia trachomatis Serovar L2 Can Utilize Exogenous Lipoic Acid through the Action of the Lipoic Acid Ligase LplA1

2010 ◽  
Vol 192 (23) ◽  
pp. 6172-6181 ◽  
Author(s):  
Aishwarya V. Ramaswamy ◽  
Anthony T. Maurelli
Keyword(s):  
Chlamydia Trachomatis ◽  
Lipoic Acid ◽  
Open Reading Frames ◽  
Sequence Motifs ◽  
Gene Encoding ◽  
E Coli ◽  
Genes Encoding ◽  
Surrogate Host ◽  
Lipoate Synthase

ABSTRACT Lipoic acid is an essential protein bound cofactor that is vital for the functioning of several important enzymes involved in central metabolism. Genomes of all sequenced chlamydiae show the presence of two genes encoding lipoic acid ligases and one gene encoding a lipoate synthase. However, the roles of these proteins in lipoic acid utilization or biosynthesis have not yet been characterized. The two distinct lipoic acid ligases in Chlamydia trachomatis serovar L2, LplA1Ct and LplA2Ct (encoded by the open reading frames ctl0537 and ctl0761) display moderate identity with Escherichia coli LplA (30 and 27%, respectively) but possess amino acid sequence motifs that are well conserved among all lipoyl protein ligases. The putative lipoic acid synthase LipACt, encoded by ctl0815, is ca. 43% identical to the E. coli LipA homolog. We demonstrate here the presence of lipoylated proteins in C. trachomatis serovar L2 and show that the lipoic acid ligase LplA1Ct is capable of utilizing exogenous lipoic acid for the lipoylation Therefore, host-derived lipoic acid may be important for intracellular growth and development. Based on genetic complementation in a surrogate host, our study also suggests that the C. trachomatis serovar L2 LipA homolog may not be functional in vivo.

Characterization of monocot and dicot plant S-adenosyl-l-methionine decarboxylase gene families including identification in the mRNA of a highly conserved pair of upstream overlapping open reading frames

2001 ◽  
Vol 353 (2) ◽  
pp. 403-409 ◽  
Author(s):  
Marina FRANCESCHETTI ◽  
Colin HANFREY ◽  
Sonia SCARAMAGLI ◽  
Patrizia TORRIGIANI ◽  
Nello BAGNI ◽  
...  
Keyword(s):  
Gene Families ◽  
Open Reading Frames ◽  
Pcr Analysis ◽  
Gene Encoding ◽  
Cdna Sequences ◽  
Regulatory Enzymes ◽  
C Terminus ◽  
Genes Encoding ◽  
Plant Genes ◽  
Reading Frames

S-Adenosyl-L-methionine decarboxylase (AdoMetDC; EC 4.1.1.50) is one of the key regulatory enzymes in the biosynthesis of polyamines. Isolation of genomic and cDNA sequences from rice and Arabidopsis had indicated that this enzyme is encoded by a small multigene family in monocot and dicot plants. Analysis of rice, maize and Arabidopsis AdoMetDC cDNA species revealed that the monocot enzyme possesses an extended C-terminus relative to dicot and human enzymes. Interestingly, we discovered that all expressed plant AdoMetDC mRNA 5´ leader sequences contain a highly conserved pair of overlapping upstream open reading frames (uORFs) that overlap by one base. The 5´ tiny uORF consists of two or three codons and the 3´ small uORF encodes 50Ő54 residues. Sequences of the small uORFs are highly conserved between monocot, dicot and gymnosperm AdoMetDC mRNA species and the C-terminus of the plant small uORFs is conserved with the C-terminus of nematode AdoMetDC uORFs; such a conserved arrangement is strongly suggestive of a translational regulatory mechanism. No introns were found in the main AdoMetDC proenzyme ORF from any of the plant genes encoding AdoMetDC, whereas introns were found in conserved positions flanking the overlapping uORFs. The absence of the furthest 3´ intron from the Arabidopsis gene encoding AdoMetDC2 suggests that this intron was lost recently. Reverse-transcriptase-mediated PCR analysis of the two Arabidopsis genes for AdoMetDC indicated that AdoMetDC1 is abundant and ubiquitous, whereas the gene for AdoMetDC2 is expressed preferentially in leaves and inflorescences. Investigation of recently released Arabidopsis genome sequences has revealed that in addition to the two genes encoding AdoMetDC isolated as part of the present work, four additional genes are present in Arabidopsis but they are probably not expressed.

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