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1.
The LexA regulated genes of the Clostridium difficile
Beata Maria Walter, Maja Rupnik, Vesna Hodnik, Gregor Anderluh, Bruno Dupuy, Nejc Paulič, Darja Žgur-Bertok, Matej Butala, 2014, izvirni znanstveni članek

Opis: Background: The SOS response including two main proteins LexA and RecA, maintains the integrity of bacterial genomes after DNA damage due to metabolic or environmental assaults. Additionally, derepression of LexA-regulated genes can result in mutations, genetic exchange and expression of virulence factors. Here we describe the first comprehensive description of the in silico LexA regulon in Clostridium difficile, an important human pathogen. Results: We grouped thirty C. difficile strains from different ribotypes and toxinotypes into three clusters according to lexA gene/protein variability. We applied in silico analysis coupled to surface plasmon resonance spectroscopy (SPR) and determined 16 LexA binding sites in C. difficile. Our data indicate that strains within the cluster, as defined by LexA variability, harbour several specific LexA regulon genes. In addition to core SOS genes: lexA, recA, ruvCA and uvrBA, we identified a LexA binding site on the pathogenicity locus (PaLoc) and in the putative promoter region of several genes involved in housekeeping, sporulation and antibiotic resistance. Conclusions: Results presented here suggest that in C. difficile LexA is not merely a regulator of the DNA damage response genes but also controls the expression of dozen genes involved in various other biological functions. Our in vitro results indicate that in C. difficile inactivation of LexA repressor depends on repressor`s dissociation from the operators. We report that the repressor`s dissociation rates from operators differentiate, thus the determined LexA-DNA dissociation constants imply on the timing of SOS gene expression in C. difficile.
Ključne besede: Clostridium difficile, antibiotic resistance, toxin regulation, SOS system, surface plasmon resonance, SPR, LexA repressor
Objavljeno v DKUM: 29.06.2017; Ogledov: 993; Prenosov: 338
.pdf Celotno besedilo (1,16 MB)
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2.
The SOS response master regulator LexA is associated with sporulation, motility and biofilm formation in Clostridium difficile
Beata Maria Walter, Stephen Cartman, Nigel Peter Minton, Matej Butala, Maja Rupnik, 2015, izvirni znanstveni članek

Opis: The LexA regulated SOS network is a bacterial response to DNA damage of metabolic or environmental origin. In Clostridium difficile, a nosocomial pathogen causing a range of intestinal diseases, the in-silico deduced LexA network included the core SOS genes involved in the DNA repair and genes involved in various other biological functions that vary among different ribotypes. Here we describe the construction and characterization of a lexA ClosTron mutant in C. difficile R20291 strain. The mutation of lexA caused inhibition of cell division resulting in a filamentous phenotype. The lexA mutant also showed decreased sporulation, a reduction in swimming motility, greater sensitivity to metronidazole, and increased biofilm formation. Changes in the regulation of toxin A, but not toxin B, were observed in the lexA mutant in the presence of sub-inhibitory concentrations of levofloxacin. C. difficile LexA is, therefore, not only a regulator of DNA damage but also controls many biological functions associated with virulence.
Ključne besede: toxins, bacterial sporulation, DNA damage, antibiotics
Objavljeno v DKUM: 19.06.2017; Ogledov: 887; Prenosov: 339
.pdf Celotno besedilo (6,06 MB)
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