scholarly journals Study of Urease gene and O-Antigen Cluster Gene of Proteus Mirabilis Isolated from Urinary Stones Patients

2017 ◽  
Vol 1 (2) ◽  
Author(s):  
Mohanad Jawad Kadhim
Keyword(s):  
Proteus Mirabilis ◽  
Urinary Stones ◽  
O Antigen ◽  
Urease Gene ◽  
Cluster Gene

Differential regulation of the Proteus mirabilis urease gene cluster by UreR and H-NS

Microbiology ◽  
2003 ◽  
Vol 149 (12) ◽  
pp. 3383-3394 ◽  
Author(s):  
Carrie A. Poore ◽  
Harry L. T. Mobley
Keyword(s):  
Urinary Tract ◽  
Proteus Mirabilis ◽  
Intergenic Region ◽  
Urinary Stones ◽  
E Coli ◽  
Urease Gene ◽  
Genes Encoding ◽  
Gel Shift Assay ◽  
Dna Fragment ◽  
Hydrolysis Of

Proteus mirabilis, a cause of catheter-associated urinary tract infection, relies on several virulence factors to colonize the urinary tract. Among these, urease contributes to the development of urinary stones resulting from the increase in local pH due to urease-mediated hydrolysis of urea to NH3 and CO2. UreR, an AraC-like transcriptional activator, activates transcription of the genes encoding the urease subunits and accessory proteins (ureDABCEFG) in the presence of urea. UreR also initiates transcription of its own gene in a urea-inducible manner by binding to the intergenic region between ureR and ureD. The intergenic region contains poly(A) tracts that appear to be the target of H-NS. It has been shown that Escherichia coli and P. mirabilis H-NS acts to repress transcription of ureR in an E. coli model system. It was hypothesized that H-NS represses urease gene expression in the absence of UreR and urea by binding to the intergenic region. To demonstrate this the P. mirabilis hns gene was cloned and the 15·6 kDa H-NS was overexpressed and purified as a myc-His tail fusion. Using a gel shift assay, purified H-NS-myc-His bound preferentially to a 609 bp DNA fragment containing the entire ureR-ureD intergenic region. H-NS and UreR were able to displace each other from the ureR-ureD intergenic region. Circular permutation analysis revealed that the intergenic region is bent. Moreover, H-NS recognizes this curvature, binds the DNA fragment and induces further bending of the DNA as shown by a circular ligation assay. The effects of H-NS, urea and temperature (25 vs 37 °C) on urease expression were shown in E. coli containing an hns knockout and P. mirabilis where expression was increased at 37 °C. Increased transcription from pureR was seen in the E. coli hns knockout when temperature was increased from 25 to 37 °C. These findings suggest H-NS and UreR differentially regulate urease in a negative and positive manner, respectively.

scholarly journals Proteus mirabilis Urease: Unsuspected Non-Enzymatic Properties Relevant to Pathogenicity

2021 ◽  
Vol 22 (13) ◽  
pp. 7205
Author(s):  
Matheus V. C. Grahl ◽  
Augusto F. Uberti ◽  
Valquiria Broll ◽  
Paula Bacaicoa-Caruso ◽  
Evelin F. Meirelles ◽  
...  
Keyword(s):  
Proteus Mirabilis ◽  
Stone Formation ◽  
Cell Types ◽  
Bioinformatic Analysis ◽  
Urinary Stone ◽  
Enzymatic Properties ◽  
Hek293 Cells ◽  
Urinary Stones ◽  
Isolated Cells ◽  
Tnf Α

Infection by Proteus mirabilis causes urinary stones and catheter incrustation due to ammonia formed by urease (PMU), one of its virulence factors. Non-enzymatic properties, such as pro-inflammatory and neurotoxic activities, were previously reported for distinct ureases, including that of the gastric pathogen Helicobacter pylori. Here, PMU was assayed on isolated cells to evaluate its non-enzymatic properties. Purified PMU (nanomolar range) was tested in human (platelets, HEK293 and SH-SY5Y) cells, and in murine microglia (BV-2). PMU promoted platelet aggregation. It did not affect cellular viability and no ammonia was detected in the cultures’ supernatants. PMU-treated HEK293 cells acquired a pro-inflammatory phenotype, producing reactive oxygen species (ROS) and cytokines IL-1β and TNF-α. SH-SY5Y cells stimulated with PMU showed high levels of intracellular Ca2+ and ROS production, but unlike BV-2 cells, SH-SY5Y did not synthesize TNF-α and IL-1β. Texas Red-labeled PMU was found in the cytoplasm and in the nucleus of all cell types. Bioinformatic analysis revealed two bipartite nuclear localization sequences in PMU. We have shown that PMU, besides urinary stone formation, can potentially contribute in other ways to pathogenesis. Our data suggest that PMU triggers pro-inflammatory effects and may affect cells beyond the renal system, indicating a possible role in extra-urinary diseases.

Structural and serological studies of the O-antigen of Proteus mirabilis O-9

2003 ◽  
Vol 338 (11) ◽  
pp. 1191-1196 ◽  
Author(s):  
Anna N. Kondakova ◽  
Rafal Fudala ◽  
Sof'ya N. Senchenkova ◽  
Aleksander S. Shashkov ◽  
Yuriy A. Knirel ◽  
...  
Keyword(s):  
Proteus Mirabilis ◽  
O Antigen ◽  
Serological Studies

Structure of the O-specific polysaccharide of Proteus mirabilis O11, another Proteus O-antigen containing an amide of d-galacturonic acid with l-threonine

1999 ◽  
Vol 323 (1-4) ◽  
pp. 81-86 ◽  
Author(s):  
Nikolay P Arbatsky ◽  
Alexander S Shashkov ◽  
Elzbieta Literacka ◽  
Göran Widmalm ◽  
Wieslaw Kaca ◽  
...  
Keyword(s):  
Proteus Mirabilis ◽  
Galacturonic Acid ◽  
O Antigen ◽  
Specific Polysaccharide

scholarly journals Structural and serological studies on the O-antigen of Proteus mirabilis O14, a new polysaccharide containing 2-[(R)-1-carboxyethylamino]ethyl phosphate

1999 ◽  
Vol 261 (2) ◽  
pp. 347-353 ◽  
Author(s):  
Andrei V. Perepelov ◽  
Elzbieta Ujazda ◽  
Sof'ya N. Senchenkova ◽  
Alexander S. Shashkov ◽  
Wieslaw Kaca ◽  
...  
Keyword(s):  
Proteus Mirabilis ◽  
O Antigen ◽  
Serological Studies ◽  
Ethyl Phosphate

scholarly journals Loss of the WaaL O-Antigen Ligase Prevents Surface Activation of the Flagellar Gene Cascade in Proteus mirabilis

2010 ◽  
Vol 192 (12) ◽  
pp. 3213-3221 ◽  
Author(s):  
Randy M. Morgenstein ◽  
Katy M. Clemmer ◽  
Philip N. Rather
Keyword(s):  
Proteus Mirabilis ◽  
Lipid A ◽  
Response Regulator ◽  
Soft Agar ◽  
Solid Surfaces ◽  
Loss Of Function ◽  
Swarmer Cell ◽  
Gram Negative ◽  
O Antigen ◽  
Enterobacterial Common Antigen

ABSTRACT Proteus mirabilis is a Gram-negative bacterium that undergoes a physical and biochemical change from a vegetative swimmer cell (a typical Gram-negative rod) to an elongated swarmer cell when grown on a solid surface. In this study, we report that a transposon insertion in the waaL gene, encoding O-antigen ligase, blocked swarming motility on solid surfaces but had little effect on swimming motility in soft agar. The waaL mutant was unable to differentiate into a swarmer cell. Differentiation was also prevented by a mutation in wzz, encoding a chain length determinant for O antigen, but not by a mutation in wzyE, encoding an enzyme that polymerizes enterobacterial common antigen, a surface polysaccharide different from the lipid A::core. In wild-type P. mirabilis, increased expression of the flhDC operon occurs after growth on solid surfaces and is required for the high-level expression of flagellin that is characteristic of swarmer cells. However, in both the waaL and the wzz mutants, the flhDC operon was not activated during growth on agar. A loss-of-function mutation in the rcsB response regulator or overexpression of flhDC restored swarming to the waaL mutant, despite the absence of O antigen. Therefore, although O antigen may serve a role in swarming by promoting wettability, the loss of O antigen blocks a regulatory pathway that links surface contact with the upregulation of flhDC expression.

scholarly journals Draft Genome of Proteus mirabilis Serogroup O18 Elaborating Phosphocholine-Decorated O Antigen

2021 ◽  
Vol 11 ◽  
Author(s):  
Grzegorz Czerwonka ◽  
Dawid Gmiter ◽  
Katarzyna Durlik-Popińska
Keyword(s):  
Proteus Mirabilis ◽  
Draft Genome ◽  
Gc Content ◽  
Gene Clusters ◽  
Gene Encoding ◽  
Protein Coding ◽  
O Antigen ◽  
Ctp:Phosphocholine Cytidylyltransferase ◽  
Annotation Server ◽  
Tract Infections

Proteus mirabilis is a pathogenic, Gram-negative, rod-shaped bacterium that causes ascending urinary tract infections. Swarming motility, urease production, biofilm formation, and the properties of its lipopolysaccharide (LPS) are all factors that contribute to the virulence of this bacterium. Uniquely, members of the O18 serogroup elaborate LPS molecules capped with O antigen polymers built of pentasaccharide repeats; these repeats are modified with a phosphocholine (ChoP) moiety attached to the proximal sugar of each O unit. Decoration of the LPS with ChoP is an important surface modification of many pathogenic and commensal bacteria. The presence of ChoP on the bacterial envelope is correlated with pathogenicity, as decoration with ChoP plays a role in bacterial adhesion to mucosal surfaces, resistance to antimicrobial peptides and sensitivity to complement-mediated killing in several species. The genome of P. mirabilis O18 is 3.98 Mb in size, containing 3,762 protein-coding sequences and an overall GC content of 38.7%. Annotation performed using the RAST Annotation Server revealed genes associated with choline phosphorylation, uptake and transfer. Moreover, amino acid sequence alignment of the translated licC gene revealed it to be homologous to LicC from Streptococcus pneumoniae encoding CTP:phosphocholine cytidylyltransferase. Recognized homologs are located in the O antigen gene clusters of Proteus species, near the wzx gene encoding the O antigen flippase, which translocates lipid-linked O units across the inner membrane. This study reveals the genes potentially engaged in LPS decoration with ChoP in P. mirabilis O18.

scholarly journals H-NS Is a Repressor of the Proteus mirabilis Urease Transcriptional Activator GeneureR

2000 ◽  
Vol 182 (9) ◽  
pp. 2649-2653 ◽  
Author(s):  
Christopher Coker ◽  
Olubunmi O. Bakare ◽  
Harry L. T. Mobley
Keyword(s):  
Nucleotide Sequence ◽  
Proteus Mirabilis ◽  
Binding Sites ◽  
Recombinant Plasmid ◽  
Urease Activity ◽  
Transcriptional Activator ◽  
Galactosidase Activity ◽  
Wild Type ◽  
Urease Gene ◽  
In Trans

ABSTRACT Expression of Proteus mirabilis urease is governed by UreR, an AraC-like positive transcriptional activator. A poly(A) tract nucleotide sequence, consisting of A6TA2CA2TGGTA5GA6TGA5, is located 16 bp upstream of the ς70-likeureR promoter P2. Since poly(A) tracts of DNA serve as binding sites for the gene repressor histone-like nucleoid structuring protein (H-NS), we measured β-galactosidase activity of wild-typeEscherichia coli MC4100 (H-NS+) and its isogenic derivative ATM121 (hns::Tn10) (H-NS−) harboring a ureR-lacZ operon fusion plasmid (pLC9801). β-Galactosidase activity in the H-NS− host strain was constitutive and sevenfold greater (P < 0.0001) than that in the H-NS+ host. A recombinant plasmid containing cloned P. mirabilis hns was able to complement and restore repression of the ureR promoter in the H-NS−host when provided in trans. Deletion of the poly(A) tract nucleotide sequence from pLC9801 resulted in an increase in β-galactosidase activity in the H-NS+ host to nearly the same levels as that observed for wild-type pLC9801 harbored by the H-NS−host. Urease activity in strains harboring the recombinant plasmid pMID1010 (encoding the entire urease gene cluster of P. mirabilis) was equivalent in both the H-NS−background and the H-NS+ background in the presence of urea but was eightfold greater (P = 0.0001) in the H-NS− background in the absence of urea. We conclude that H-NS represses ureR expression in the absence of urea induction.

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