- Volume 146, Issue 3, 2000
Volume 146, Issue 3, 2000
- Sgm Special Lecture
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- Microbiology Comment
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- Biochemistry
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Proteins complexed to the P1 adhesin of Mycoplasma pneumoniae
More LessAdherence of Mycoplasma pneumoniae to host cells requires several mycoplasmal membrane proteins and cytoskeleton-like proteins in addition to the adhesin P1, a transmembrane protein of 170 kDa. To analyse interactions of the P1 adhesin with other membrane proteins or with cytoskeleton-like proteins, cross-linking studies were performed in vivo using the permeant reagent paraformaldehyde. The cross-linked protein complex was isolated by immunoaffinity chromatography, and proteins complexed to the P1 protein were identified by immunoblot analysis followed by high mass accuracy tryptic peptide mapping using matrix-assisted laser desorption/ionization mass spectrometry (MALDI MS). In addition to the P1 protein and a truncated form of the same protein, the adhesin-related 30 kDa protein, two membrane proteins of 40 and 90 kDa, the cytoskeleton-associated 65 kDa protein and two cytoskeleton-forming proteins, HMW1 and HMW3, were found to be components of the isolated protein complex. Furthermore, the cross-linked complex contained the chaperone DnaK and the E1α subunit of pyruvate dehydrogenase. In summary, it was shown that cytadherence-associated membrane proteins are located in close proximity to cytoskeleton-like proteins, suggesting a functional interaction between membrane and cytoskeleton-like proteins. DnaK might be involved in translocation of proteins from the cytoplasm to the membrane and pyruvate dehydrogenase might be a structural protein of the attachment organelle.
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- Bioenergetics And Transport
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Protonmotive force regulates the membrane conductance of Streptococcus bovis in a non-ohmic fashion
More LessBecause the DCCD (dicyclohexylcarbodiimide)-sensitive, F-ATPase-mediated, futile ATP hydrolysis of non-growing Streptococcus bovis JB1 cells was not affected by sodium or potassium, ATP hydrolysis appeared to be dependent only upon the rate of proton flux across the cell membrane. However, available estimates of bacterial proton conductance were too low to account for the rate of ATP turnover observed in S. bovis. When de-energized cells were subjected to large pH gradients (2·75 units, or −170 mV), internal pH declined at a rate of 0·15 pH units s−1. Based on an estimated cellular buffering capacity of 200 nmol H+ (mg protein)−1 per pH unit, H+ flux across the cell membrane (at −170 mV) was 108 mmol (g protein)−1 h−1. When potassium-loaded cells were treated with valinomycin in low-potassium buffers, initial K+ efflux generated membrane potentials in close agreement with values predicted by the Nernst equation. These artificial membrane potentials drove H+ uptake, and H+ influx was counterbalanced by a further loss of cellular K+. Flame photometry indicated that the rate of K+ loss was 215 (±26) mmol K+ (g protein)−1 h−1 at −170 mV, but the potassium-sensitive fluorescent compound CD222 indicated that this rate was only 110 (±44) mmol K+ (g protein)−1 h−1. As pH gradients or membrane potentials were reduced, the rate of H+ flux declined in a non-ohmic fashion, and all rates were <25 mmol (g protein)−1 h−1 at a driving force of −80 mV. Previous estimates of bacterial proton flux were based on low and unphysiological protonmotive forces, and the assumption that H+ influx rate would be ohmic. Rates of H+ influx into S. bovis cells [as high as 9×10−11 mol H+ (cm membrane)−2 s−1] were similar to rates reported for respiring mitochondria, but were at least 20-fold greater than any rate previously reported in lactic acid bacteria.
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l–[U–14C]Lactate binding to a 43 kDa protein in plasma membranes of Candida utilis
More LessTo identify the putative lactate transporter protein of Candida utilis, plasma membranes from cells grown either on lactic acid (presence of lactate proton symport) or glucose (absence of lactate proton symport) were incubated with l–[U–14C]lactic acid and the membrane proteins were then separated by SDS-PAGE. A well-defined peak of radioactivity occurred in the lane of the gel containing plasma membrane proteins from lactic-acid-grown cells but not from glucose-grown cells. Binding was inhibited by unlabelled pyruvate and lactate, whereas succinate and citrate were not inhibitory. The monocarboxylate transporter inhibitor of animal cells, 4,4′-diisothiocyanato-stilbene-2,2′-disulfonate, competitively inhibited the lactate proton symport in the whole yeast and also inhibited lactate binding to proteins of isolated plasma membranes. The polypeptide pattern of plasma membranes from lactic-acid-grown cells revealed a 43 kDa polypeptide associated with the peak of labelled lactate. Altogether the results suggest that this polypeptide is either the lactate transporter or a component of it.
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- Environmental Microbiology
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Yeast spores seem to be involved in biological phosphate removal: a microscopic in situ case study
More LessThe principal polyphosphate-accumulating organism (PAO) in a biological-phosphate-removal activated-sludge process was assessed microscopically. The organism was recognized by its distinct morphotype most easily after polyphosphate staining. The PAO occurred in large, homogeneous clusters. The cells of the PAO were the biggest cells abounding in the sludge – clearly bigger than average sludge bacteria. Typical of the principal PAO was a variation of cell size, even in fresh sludge. In acetate minimal medium containing ampicillin, the original principal PAO clusters were converted to clusters of clearly larger, polyphosphate-containing, vegetative yeast-like cells. Cycloheximide addition inhibited this and caused flock disintegration, disappearance of the principal PAO clusters and growth of free bacteria. The cell wall of the principal PAO was not of the usual bacterial character. It showed anomalous Gram staining, stained for chitin (not found in bacteria) and bound concanavalin A, like cell walls of many yeasts. In addition, the PAO cell wall was resistant to lysozyme, but sensitive to an enzyme mixture that lyses yeast cell walls. It was concluded that the principal PAO cells in the studied sludge were clustered spores of a yeast.
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Occurrence of natural dixenic associations between the symbiont Photorhabdus luminescens and bacteria related to Ochrobactrum spp. in tropical entomopathogenic Heterorhabditis spp. (Nematoda, Rhabditida)
More LessThe EMBL accession numbers for the 16S rDNA sequences reported in this paper are AJ245941 (PR17/sat), AJ249458 (FRG11/sat) and AJ249459 (DO23/sat).
Bacteria naturally associated with the symbiont Photorhabdus luminescens subsp. akhurstii were isolated from the entomopathogenic nematode Heterorhabditis indica. Bacterial isolates distinct from P. luminescens subsp. akhurstii were obtained from 33% of the samples. Fourteen bacterial isolates, from nematodes collected from three different Caribbean islands, were characterized by conventional phenotypic tests, restriction fragment length polymorphism and sequence analyses of PCR-amplified 16S rRNA genes (16S rDNAs). Isolates were grouped into three genotypes, each one being associated with one Caribbean island. Phenotypic characteristics and 16S rDNA analysis showed that the Photorhabdus-associated bacteria were closely related to Ochrobactrum anthropi for the group from Guadeloupe, and to Ochrobactrum intermedium for the two groups from the Dominican Republic and Puerto Rico. No pathogenicity of the Ochrobactrum spp. to the insects Galleria mellonella and Spodoptera littoralis (Lepidoptera) was detected. Since Ochrobactrum spp. are considered as human opportunist pathogens, the mass production of entomopathogenic nematodes for biological control requires strict vigilance.
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- Genetics And Molecular Biology
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Stability by multimer resolution of pJHCMW1 is due to the Tn1331 resolvase and not to the Escherichia coli Xer system
More LessThe GenBank accession number for the sequence of the pJHCMW1 EcoRI–SacI fragment reported in this paper is AF135798.
The plasmid pJHCMW1 encodes resistance to several aminoglycosides and β-lactams and consists of a copy of the transposon Tn1331, a region including the replication functions, and a sequence with homology to ColE1 cer, designated mwr. In this work, the role of this cer-like site in ensuring the stable inheritance of pJHCMW1 by multimer resolution was studied. The Escherichia coli Xer site-specific recombination system acts at sites such as ColE1 cer to resolve plasmid multimers formed by homologous recombination, thereby maintaining plasmids in a monomeric state and helping to ensure stable plasmid inheritance. Despite its high similarity to ColE1 cer, the pJHCMW1 mwr was a poor substrate for Xer recombination in E. coli and did not contribute significantly to plasmid stability. Instead, the Tn1331 co-integrate resolution system was highly active at resolving pJHCMW1 multimers and ensured the stable inheritance of pJHCMW1. Although Xer recombination at pJHCMW1 mwr was inefficient in E. coli, the recombination that did occur was dependent on ArgR, PepA, XerC and XerD. A supercoiled circular DNA molecule containing two pJHCMW1 mwr sites in direct repeat yielded Holliday-junction-containing product when incubated with ArgR, PepA, XerC and XerD in vitro, confirming that pJHCMW1 mwr is a functional recombination site. However, unlike cer, some Holliday-junction-containing product could be detected for mwr in the absence of ArgR, although addition of this protein resulted in formation of more Holliday junctions. Binding experiments demonstrated that XerD bound to pJHCMW1 mwr core with a high affinity, but that XerC bound to this site very poorly, even in the presence of XerD.
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Mu DNA reintegration upon excision: evidence for a possible involvement of nucleoid folding
More LessMutations induced by the integration of a Mugem2ts prophage can revert at frequencies around 1×10−6. In these revertant clones, the prophage excised from its original localization is not lost but reintegrated elsewhere in the host genome. One of the most intriguing aspects of this process is that the prophage reintegration is not randomly distributed: there is a strong correlation between the original site of insertion (the donor site) and the target site of the phage DNA migration (the receptor site). In this paper, it is shown that in the excision–reintegration process mediated by Mugem2ts, the position of the initial prophage site strongly influences the location of the reintegration site. In addition, for each donor site, the receptor site is a discrete DNA region within which the excised Mu DNA can reintegrate and the two sites implicated in phage DNA migration must be located on the same DNA molecule. These data suggest the involvement of nucleoid folding in the excision–reintegration process.
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A bacteriophage-like particle from Bartonella bacilliformis
More LessBartonella bacilliformis and Bartonella henselae, the respective agents of Oroya fever and cat-scratch disease in humans, are known to produce bacteriophage-like particles (BLPs) that package 14 kbp segments of the host chromosome. Data from this study suggest that other Bartonella species including Bartonella quintana, Bartonella doshiae and Bartonella grahamii also contain similar BLPs, as evidenced by the presence of a 14 kbp extrachromosomal DNA element in their genomes, whereas Bartonella elizabethae and Bartonella clarridgeiae do not. A purification scheme utilizing chloroform, DNase I and centrifugation was devised to isolate BLPs from B. bacilliformis. Intact BLPs were observed by transmission electron microscopy and were round to icosahedral in shape and approximately 80 nm in diameter. RFLP and Southern blot analysis of BLP DNA from B. bacilliformis suggest that packaging, while non-selective, is less than the near-random packaging previously reported for the B. henselae phage. Data also suggest that the linear, double-stranded BLP DNA molecules have blunt ends with non-covalently closed termini. Packaging of the BLP DNA molecules into a protein coat appears to be closely related to nucleic acid synthesis, as unpackaged phage DNA is not detectable within the host cell. SDS-PAGE analysis of purified BLPs from B. bacilliformis showed three major proteins with apparent molecular masses of 32, 34 and 36 kDa; values that closely correspond to proteins found in B. henselae BLPs. Western blot analysis performed with patient convalescent serum showed that BLP proteins are slightly immunogenic in humans. To determine if BLPs contribute to horizontal gene transfer, mutants of B. bacilliformis were generated by allelic exchange with an internal fragment of the 16S–23S rDNA intergenic spacer region and a suicide vector construct, termed pKB1. BLPs from one of the resultant strains were able to package the mutagenized region containing the kanamycin-resistance cassette; however, numerous approaches and attempts at intraspecies transduction using these BLPs were unsuccessful.
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Identification of a gene cluster for antibacterial polyketide-derived antibiotic biosynthesis in the nystatin producer Streptomyces noursei ATCC 11455
More LessThe GenBank accession numbers for the sequences reported in this paper are AF071512 for ORF1, AF071513 for ORF2, AF071514 for ORF3, AF071515 for ORF4, AF071516 for ORF5, AF071517 for ORF6, AF071518 for ORF7, AF071519 for gdhA, AF071520 for ORF8, AF071521 for ORF9, AF071522 for ORF10 and AF071523 for ORF11.
Streptomyces noursei ATCC 11455 produces the antifungal polyene antibiotic nystatin containing the deoxysugar moiety mycosamine. Part of the deoxythymidyl diphosphate (TDP)-glucose dehydratase gene (gdhA) known to be involved in deoxysugar biosynthesis was amplified by PCR from genomic DNA of S. noursei ATCC 11455. A gene library for S. noursei was made and screened with the gdhA probe. Several overlapping phage clones covering about 30 kb of the S. noursei genome were physically mapped. A partial DNA sequencing analysis of this region resulted in the identification of several putative genes typical of macrolide antibiotic biosynthetic gene clusters. A gene-transfer system for S. noursei has been established, and gene deletion or disruption experiments within the putative biosynthetic gene cluster were performed. All of the knock-out mutants retained the ability to produce nystatin, suggesting that the identified gene cluster is not involved in biosynthesis of this antibiotic. Culture extracts from the wild-type strain and three knock-out mutants were analysed by TLC followed by a bioassay against Micrococcus luteus. Two antibacterial compounds were found to be synthesized by the wild-type strain while only one was produced by the mutants. This provided evidence for the involvement of the identified gene cluster in the biosynthesis of a presumably novel antibacterial macrolide antibiotic in S. noursei.
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Characterization of the genetic locus responsible for production and immunity of carnobacteriocin A: the immunity gene confers cross-protection to enterocin B
More LessThe GenBank/EMBL/DDBJ accession number for the sequence reported in this paper is AF207838
Carnobacteriocin A (CbnA) is a regulated bacteriocin produced by Carnobacterium piscicola LV17A that is encoded on a 72 kb plasmid. A 10·0 kb fragment from this plasmid that contained information necessary for bacteriocin production and immunity was cloned and sequenced. Genetic analysis showed the presence of the previously sequenced structural gene for CbnA, as well as genes encoding proteins homologous to dedicated bacteriocin transport proteins and proteins of three-component signal transduction systems. The induction factor (CbnX) was chemically synthesized and induced CbnA production at 10−11 M or higher in a C. piscicola LV17A culture that had lost the ability to produce bacteriocin as a result of dilution. The gene cbiA for the immunity protein is not located in typical close proximity to the structural gene for CbnA and is encoded in the opposite orientation. CbiA has homology with EniB, the immunity protein for enterocin B that is also encoded in the opposite orientation to the bacteriocin gene. CbiA and EniB cross-protected against the corresponding bacteriocins.
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Polyadenylylation in mycobacteria: evidence for oligo(dT)-primed cDNA synthesis
More LessThe ability of mRNA to direct synthesis of cDNA in the presence of oligo(dT) was analysed using a novel application of fluorescein-11-dUTP incorporation into cDNA by reverse transcriptase. Evidence is provided for the first time that a majority of the mycobacterial mRNA pool is polyadenylylated. mRNA transcripts of hsp65 were also amplified with specific primers from the oligo(dT)-primed cDNA preparation in Mycobacterium bovis BCG, M. smegmatis and M. vaccae. Furthermore, PCR amplication of cDNAs for genes entD, entC and trpE2 from M. bovis BCG yielded the expected products when reverse transcription was primed with oligo(dT), suggesting that polyadenylylation is a general phenomenon in mycobacteria.
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Overproduction of the secretin OutD suppresses the secretion defect of an Erwinia chrysanthemi outB mutant
More LessOutB is a component of the Erwinia chrysanthemi Out secretion machinery. Homologues of OutB have been described in two other bacteria, Klebsiella oxytoca and Aeromonas hydrophila, but their requirement in the secretion process seems to be different. Study of OutB topology with the BlaM topology probe suggests that it is an inner-membrane protein with a large periplasmic domain. However, fractionation experiments indicate that it could be associated with the outer membrane through its C-terminal part. The secretion deficiency of an Erw. chrysanthemi outB mutant can be reversed by the addition of an inducer of the kdgR regulon. It was shown that this effect results from the increased expression of the secretin OutD and that secretion can be restored in an outB mutant by introducing the outD gene on a plasmid. Several experiments suggest an interaction between OutB and OutD. In Erw. chrysanthemi, the presence of OutD stabilizes OutB. OutD expressed in Escherichia coli can be protected from proteolytic degradation by the coexpression of OutB. This effect does not require the N-terminal, transmembrane segment of outB. OutB can be cross-linked with OutD by formaldehyde. These results indicate that OutB could act with OutD in the functioning of the Out secretion machinery.
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The gene pvaB encodes oxidized polyvinyl alcohol hydrolase of Pseudomonas sp. strain VM15C and forms an operon with the polyvinyl alcohol dehydrogenase gene pvaA
More LessThe DDBJ accession number for the sequence reported in this paper is AB008494.
A 5·7 kbp SphI fragment containing the polyvinyl alcohol (PVA) dehydrogenase gene pvaA and its 1·9 kbp 5′-flanking region was cloned from the PVA-degrading bacterium Pseudomonas sp. VM15C. The pvaB gene, encoding oxidized PVA hydrolase, was found in the region upstream of pvaA. Sequence data and expression studies indicated that pvaA and B constitute an operon in the order pvaBA. The pvaB gene encoded a protein of 379 amino acid residues (40610 Da), and a lipoprotein signal sequence and the lipase consensus sequence, Gly-X-Ser-X-Gly, characteristic of the active-site serine region in serine hydrolases, were detected in the deduced amino acid sequence. The pvaB product with the pvaA product constituted an enzyme system for the cleavage of PVA molecules. The pvaA product introduced β-diketone groups into the PVA molecule, and the pvaB product hydrolysed these β-diketone groups in oxidized PVA. The pvaB product also hydrolysed 4,6-nonanedione at a low rate, but not acetylacetone or 5-nonanone. It was completely inhibited by PMSF and was concluded to be a serine hydrolase. There were no proteins showing high similarity to the pvaB product in the databases, but minor similarity to a number of serine hydrolases including polyhydroxyalkanoate depolymerases was apparent.
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Molecular characterization of the ferric-uptake regulator, Fur, from Staphylococcus aureus
More LessThe GenBank accession numbers for the S. aureus fur gene and fhu operon reported in this paper are AF118839 and AF132117, respectively.
Iron is an essential nutrient for the survival and pathogenesis of bacteria, but relatively little is known regarding its transport and regulation in staphylococci. Based on the known sequences of ferric-uptake regulatory (fur) genes from several Gram-positive and Gram-negative bacteria, a fragment containing the fur homologue was cloned from a genomic library of Staphylococcus aureus RN450. Nucleotide sequence analysis of this fragment revealed the presence of a 447 bp ORF that encodes a putative 149 aa polypeptide with an apparent molecular mass of 17 kDa. A putative ferrichrome-uptake (fhu) operon, containing the conserved Fur-binding sequences (Fur box) in the promoter region, was also cloned from the same S. aureus library. To characterize the impact of Fur on the fhu operon, fur was cloned, overexpressed as a His-tagged protein and purified by Ni2+-affinity column chromatography. The recombinant protein was digested with enterokinase to remove the His tag. Electrophoretic mobility-shift assays indicated that Fur binds to the promoter region of the fhu operon in the presence of divalent cations. Fur also interacted with the promoter region of the recently reported sir operon that has been proposed to constitute a siderophore-transport system in S. aureus. The DNase I-protection assay revealed that Fur specifically binds to the Fur box located in the promoter region of the fhu operon. The primer-extension reaction indicated that the transcription-start site of the fhu operon was located inside the Fur box. S. aureus fur partially complemented a fur − mutation in Bacillus subtilis. The data suggest that Fur regulates iron-transport processes in S. aureus.
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Fur-independent regulation of iron metabolism by Irr in Bradyrhizobium japonicum
More LessBradyrhizobium japonicum expresses both Fur and Irr, proteins that mediate iron-dependent regulation of gene expression. Control of irr mRNA accumulation by iron was aberrant in a fur mutant strain, and Fur repressed an irr::lacZ promoter fusion in the presence of iron. Furthermore, metal-dependent binding of Fur to an irr gene promoter was demonstrated in a region with no significant similarity to the Fur-binding consensus DNA element. These data suggest that the modest control of irr transcription by iron is mediated by Fur. However, Irr protein levels were regulated normally by iron in the fur strain, indicating that Fur is not required for post-transcriptional control of the irr gene. Accordingly, regulation of hemB, a haem biosynthesis gene regulated by Irr, was controlled normally by iron in a fur strain. In addition, the hemA gene was shown to be controlled by Fur, but not by Irr. It was concluded that Fur cannot be the only protein by which B. japonicum cells sense and respond to iron, and that Irr may be involved in Fur-independent signal transduction. Furthermore, iron-dependent regulation of haem biosynthesis involves both Irr and Fur.
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The gene encoding IIABL Man in Streptococcus salivarius is part of a tetracistronic operon encoding a phosphoenolpyruvate:mannose/glucose phosphotransferase system
More LessThe GenBank accession number for the sequence reported in this paper is AF130465.
Glucose and mannose are transported in streptococci by the mannose-PTS (phosphoenolpyruvate:mannose phosphotransferase system), which consists of a cytoplasmic IIAB protein, called IIABMan, and an uncharacterized membrane permease. This paper reports the characterization of the man operon encoding the specific components of the mannose-PTS of Streptococcus salivarius. The man operon was composed of four genes, manL, manM, manN and manO. These genes were transcribed from a canonical promoter (Pman) into a 3·6 kb polycistronic mRNA that contained a 5′-UTR (untranslated region). The predicted manL gene product encoded a 35·5 kDa protein and contained the amino acid sequences of the IIA and IIB phosphorylation sites already determined from purified S. salivarius . Expression of manL in Escherichia coli generated a 35 kDa protein that reacted with antibodies. The predicted ManM protein had an estimated size of 27·2 kDa. ManM had similarity with IIC domains of the mannose-EII family, but did not possess the signature proposed for mannose-IIC proteins from Gram-negative bacteria. From multiple alignment analyses of sequences available in current databases, the following modified IICMan signature is proposed: GX3G[DNH]X3G[LIVM]2XG2[STL][LT][EQ]. The deduced product of manN was a hydrophobic protein with a predicted molecular mass of 33·4 kDa. The ManN protein contained an amino acid sequence similar to the signature sequence of the IID domains of the mannose-EII family. manO encoded a 13·7 kDa protein. This gene was also transcribed as a monocistronic mRNA from a promoter located in the manN–manO intergenic region. A search of current databases revealed the presence of , ManM, ManN and ManO orthologues in Streptococcus mutans, Streptococcus pyogenes, Streptococcus pneumoniae and Enterococcus faecalis. This work has elucidated the molecular structure of the mannose PTS in streptococci and enterococci, and demonstrated the presence of a putative regulatory protein (ManO) within the man operon.
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Evidence for specificity in type 4 pilus biogenesis by enteropathogenic Escherichia coli
More LessType 4 fimbriae (pili) are surface appendages that are expressed by many species of Gram-negative bacteria. Previous studies have demonstrated that Pseudomonas aeruginosa can express and assemble pilin subunits from several unrelated species, indicating a common mechanism for biogenesis of type 4 pili whereby structural subunits from one system may be interchanged with those of another. In this study, an isogenic mutant of enteropathogenic Escherichia coli (EPEC) was constructed containing the entire tcpA gene from Vibrio cholerae O395, which encodes the major structural subunit of the toxin-coregulated pilus (TCP), in place of bfpA, which encodes the major structural subunit of the bundle-forming pilus (BFP). Surprisingly, expression of type 4 pilin structures and the associated phenotype of bacterial autoaggregation in culture media were not observed for cells of the EPEC strain containing tcpA nor for those containing an additional mutation in bfpF, which otherwise is associated with a hyperfimbriate phenotype. In addition, cells of a bfpA mutant EPEC strain containing plasmids designed to express either of two different chimeric type 4 pilin subunits containing segments of BfpA and TcpA also failed to form bacterial aggregates and express type 4 pilin structures. Collectively, these results indicate that the type 4 pilin assembly system of EPEC exhibits specificity with regard to pilin subunit recognition and assembly.
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- Genomics
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Systematic study of gene expression and transcription organization in the gntZ–ywaA region of the Bacillus subtilis genome
Within the framework of the international project ‘The functional analysis of the Bacillus subtilis genome’ in Japan and Europe, the gene expression and transcription organization of the gntZ–ywaA region (160 kb) of the B. subtilis genome has been systematically analysed. First, all unanalysed genes comprising more than 80 amino acids (125 genes) in this region were inactivated through integration of plasmid pMUTIN. No essential gene was found which could not be inactivated. All the integrants grew normally in both nutrient sporulation medium and glucose minimal medium. But an integrant in the yxbG gene exhibited an oligosporogenic phenotype in the nutrient sporulation medium. The synthesis of β-galactosidase was examined, as a reporter for expression of the inactivated genes, during growth and sporulation in the two media. The results indicated that 36% of the promoters were inactive when cells were grown in at least one of these two media. Furthermore, the transcription of the 119 genes in this region was analysed by Northern blotting, resulting in a transcription map. The results indicate that the gntZ–ywaA region contains at least 24 polycistronic operons, including several published ones. The operons newly found in this work are yxaAB, yxaGH, yxaJKL, yxbBA–yxnB–asnH–yxaM, yxbCD, yxcED, yxdJK, yxeFGH, yxeKLMNOPQ, yxeR–yxxB, hutPHUIGM, bglPH–yxiE, wapA–yxxG, yxiM–deaD, katB–yxiS, yxjCDEF, yxjJI and yxkF–mmsX.
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Volume 20 (1959)
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Volume 19 (1958)
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Volume 18 (1958)
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Volume 17 (1957)
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Volume 16 (1957)
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Volume 15 (1956)
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Volume 14 (1956)
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Volume 13 (1955)
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Volume 12 (1955)
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Volume 11 (1954)
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Volume 10 (1954)
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Volume 9 (1953)
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Volume 8 (1953)
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Volume 7 (1952)
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Volume 6 (1952)
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Volume 5 (1951)
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Volume 4 (1950)
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Volume 3 (1949)
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Volume 2 (1948)
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Volume 1 (1947)