ABSTRACT The par genes of Pseudomonas aeruginosa have been studied to increase the understanding their mechanism action and role in bacterial cell. Key properties ParB protein identified are associated with different parts protein. ParB- interaction domain was mapped vivo vitro C-terminal 56 amino acids (aa); 7 aa at C terminus play an important role. dimerization P. is interchangeable KorB from plasmid RK2 (IncP1 group). part also involved ParB-ParA interactions. Purified binds specifically...

Summary The kilB locus (which is unclonable in the absence of korB ) broad‐host‐range plasmid RK2 (60kb) lies between trfA operon (co‐ordinates 16.4 to 18.2kb), which encodes a protein essential for vegetative replication, and Tra2 block conjugative transfer genes 20.0 27.0 kb). Promoter probe studies indicated that transcribed clockwise from region containing closely spaced divergent promoters, one promoter. repression both promoters by suggested may also play role stable maintenance RK2....

ParB protein of Pseudomonas aeruginosa belongs to a widely represented family chromosomally and plasmid-encoded partitioning type IA proteins. Ten putative parS sites are dispersed in the P. chromosome, with eight them localizing oriC domain. After binding parS, spreads on DNA, causing transcriptional silencing nearby genes (A. A. Bartosik et al., J. Bacteriol. 186:6983-6998, 2004). We have studied derivatives impaired spreading either due loss DNA-binding ability or oligomerization. defined...

Nucleotide sequences of the cysB region Salmonella typhimurium and Escherichia coli have been determined compared.A total 1759 nucleotides were sequenced in S. 1840 E. coli.

The parA and parB genes of Pseudomonas aeruginosa are located approximately 8 kb anticlockwise from oriC. ParA is a cytosolic protein present at level around 600 molecules per cell in exponential phase, but the drops about fivefold stationary phase. Overproduction full-length or N-terminal 85 amino acids severely inhibits growth P. putida. Both inactivation overexpression trans also lead to accumulation anucleate cells changes motility. Inactivation increases turnover rate (degradation)...

Broad-host-range IncP plasmid RK2 possesses a series of operons involved in maintenance, whose expression is coordinated by number regulators, most which are encoded the central regulatory korA-korB operon. The nucleotide sequence two new cistrons this operon, comprising what we have previously designated korF locus located between coordinates 57.0 and 56.0 kb on genome alpha RK2, presented. encode polypeptides 173 175 amino acids. Each can repress transcription from promoters for kfrA (a...

Deletions leading to complete or partial removal of ParB were introduced into the Pseudomonas aeruginosa chromosome. Fluorescence microscopy fixed cells showed that mutants lacking C-terminal domain HTH motif formed multiple, less intense foci scattered irregularly, in contrast one four per cell symmetrically distributed wild-type P. aeruginosa. All parB mutations affected both bacterial growth and swarming swimming motilities, increased production anucleate cells. Similar effects observed...

ParA and ParB homologs are involved in accurate chromosome segregation bacteria. ParBs participate the separation of ori domains by binding to parS palindromes, mainly localized close oriC. In Pseudomonas aeruginosa neither deficiency nor modification all 10 parSs is lethal. However, such mutants show not only defects but also growth retardation motility dysfunctions. Moreover, a lack parB alters expression over 1000 genes, suggesting that could interact with outside its canonical targets....

IncU plasmids are a distinctive group of mobile elements with highly conserved backbone functions and variable antibiotic resistance gene cassettes. The archetype is conjugative plasmid RA3, whose sequence (45,909 bp) shows it to be mosaic, modular replicon class I integron different from that other replicons. Functional analysis demonstrated RA3 possesses broad host range can efficiently self-transfer, replicate, maintained stably in alpha-, beta-, gammaproteobacteria. contains 50 open...

Accurate chromosome segregation to progeny cells is a fundamental process ensuring proper inheritance of genetic material. In bacteria with simple cell cycle, follows replication initiation since duplicated oriC domains start segregating opposite halves the soon after they are made. ParA and ParB proteins together specific DNA sequences parts machinery. in Pseudomonas aeruginosa important for optimal growth, nucleoid segregation, division motility. Comparative transcriptome analysis parAnull...

The trb operon of broad-host-range plasmid RK2 encodes most the genes required for formation mating-pair apparatus and is thus essential promiscuous spread this plasmid. Only two promoters, lying upstream trbA trbB, have been identified operon. trbB a protein belonging to large family proteins which function in assembly apparatuses associated with cell surface. repressor protein, one whose targets promoter. trbAp arranged as face-to-face divergent promoter trfAp, strongest three promoters...

Previous deletion and complementation analysis has indicated that the region between trfA kilBI (trbB) encodes trans-acting factor, designated trbA, required for conjugative transfer of broad host range plasmid RK2. In analysing nucleotide sequence this we have discovered a gene encoding 12 kDa polypeptide. The predicted amino acid protein shows similarity at its C-terminal to KorA from central control operon RK2 N-terminal immunity repressor phage phi 105 Bacillus subtilis as well Sin B....

Strains of escherichia coli were constructed in which the lacZ gene is fused to cysB, positive regulator cysteine regulon. The fusion strains used study regulation cysB by assaying product. introduction a allele, either on plasmid or an episome strains, resulted decrease beta-galactosidase activity. This implies that expression autoregulated its own direction transcription was determined be clockwise.

Among the mechanisms that control chromosome segregation in bacteria are highly-conserved partitioning systems comprising three components: ParA protein (a deviant Walker-type ATPase), ParB DNA-binding element) and multiple cis-acting palindromic centromere-like sequences, designated parS. Ten putative parS sites have been identified P. aeruginosa PAO1 genome, four localized close proximity of oriC six, diverged by more than one nucleotide from a perfect sequence, dispersed along chromosome....

The Tra1 region of broad host range IncPα plasmid RK2 encodes proteins essential for its promiscuous conjugative transfer and includes oriT, the site at which nicking occurs to initiate replication. Unregulated expression genes would be likely place a major burden on host. To investigate control these three transcriptional promoters from this were cloned by PCR inserted into xylEpromoter probe vectors. strength traJpand traKpwas estimated six eightfold less than strong trfApromoter is...

The trfA and trb operons, encoding genes essential for replication conjugative transfer of broad host range plasmid RK2, are transcribed divergently. Deletion analysis presented here indicates that trfAp trbAp arranged as face to promoters. presence the korA gene, whose product is known repress seven operons on including operon, shown stimulate trbAp. effect mimicked by trfAp-1 promoter down mutation, suggesting a reduction in activity required derepression activity. mutation reduces RNA...

A total of 30 kilobases DNA comprising the cysAMK region S. typhimurium was cloned as a series fragments in phage lambda 1059. The genetic organization this established through studies gene expression from subcloned pBR322 and blot hydridization analyses restriction sites chromosomal multisite deletion strains. results give order cysA-cysM-crr-ptsl-ptsH-cysK over distance approximately 12 kilobases. cysM cysA have been expressed pBR322; attempts to obtain stable derivatives carrying cysK...

Pseudomonas aeruginosa is a cause of nosocomial infections, especially in patients with cystic fibrosis and burn wounds. PAO1 strain its derivatives are widely used to study the biology this bacterium, however recent studies demonstrated differences genomes phenotypes from different laboratories.Here we report genome sequence P. PAO1161 laboratory strain, leu-, RifR, restriction-modification defective derivative, described as host IncP-8 plasmid FP2, conferring resistance mercury. Comparison...

In the majority of bacterial species, tripartite ParAB-parS system, composed an ATPase (ParA), a DNA-binding protein (ParB), and its target parS sequence(s), assists in chromosome partitioning. ParB forms large nucleoprotein complexes at parS(s), located vicinity origin chromosomal replication (oriC), which after are subsequently positioned by ParA cell poles. Remarkably, participate not only segregation but through interactions with various cellular partners they also involved other...