A multiresistance megaplasmid pLG1 bearing a hylEfm genomic island in hospital Enterococcus faecium isolates

Gomez, Jenny A. Laverde; Schaik, Willem van; Freitas, Ana R.; Coque, Teresa M.; Weaver, Keith E.; Francia, María Victoria; Witte, Wolfgang; Werner, Guido

doi:10.1016/j.ijmm.2010.08.015

Cited by 71 publications

(56 citation statements)

References 56 publications

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“…[34]. PCR for the rep pLG1 [9] was performed with primers designed previously [35]. The presence of plasmid addiction systems, relaxase genes, and the intA integrase gene of integrative and conjugative element ICE Efm1 was also verified by PCR [36–38].…”

Section: Methodsmentioning

confidence: 99%

Molecular Analysis of VanA Outbreak ofEnterococcus faeciumin Two Warsaw Hospitals: The Importance of Mobile Genetic Elements

Wardal

Markowska

Żabicka

et al. 2014

BioMed Research International

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Vancomycin-resistant Enterococcus faecium represents a growing threat in hospital-acquired infections. Two outbreaks of this pathogen from neighboring Warsaw hospitals have been analyzed in this study. Pulsed-field gel electrophoresis (PFGE) of SmaI-digested DNA, multilocus VNTR analysis (MLVA), and multilocus sequence typing (MLST) revealed a clonal variability of isolates which belonged to three main lineages (17, 18, and 78) of nosocomial E. faecium. All isolates were multidrug resistant and carried several resistance, virulence, and plasmid-specific genes. Almost all isolates shared the same variant of Tn1546 transposon, characterized by the presence of insertion sequence ISEf1 and a point mutation in the vanA gene. In the majority of cases, this transposon was located on 50 kb or 100 kb pRUM-related plasmids, which lacked, however, the axe-txe toxin-antitoxin genes. 100 kb plasmid was easily transferred by conjugation and was found in various clonal backgrounds in both institutions, while 50 kb plasmid was not transferable and occurred solely in MT159/ST78 strains that disseminated clonally in one institution. Although molecular data indicated the spread of VRE between two institutions or a potential common source of this alert pathogen, epidemiological investigations did not reveal the possible route by which outbreak strains disseminated.

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Section: Methodsmentioning

confidence: 99%

Molecular Analysis of VanA Outbreak ofEnterococcus faeciumin Two Warsaw Hospitals: The Importance of Mobile Genetic Elements

Wardal

Markowska

Żabicka

et al. 2014

BioMed Research International

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“…CwlT ICEBs1 possesses a C-terminal CHAP (cysteine-, histidine-dependent amidohydrolase/peptidase) domain with DL-endopepti-dase activity and an N-terminal domain structurally similar to lytic transglycosylases but with N-acetylmuramidase activity (18). Other members of this two-domain hydrolase family include Clostridium perfringens TcpG pCW3 (19), Streptococcus agalactiae TraG pIP501 (20), Enterococcus faecium Orf0032 pLG1 (21), and Orf14, encoded by Tn916/Tn925 ICEs (22,23). These proteins carry CHAP domains with a predicted endopeptidase or amidase activity and SLT-or FlgJ-like domains with predicted glycosylase or muramidase activities (see Fig.…”

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confidence: 99%

PrgK, a Multidomain Peptidoglycan Hydrolase, Is Essential for Conjugative Transfer of the Pheromone-Responsive Plasmid pCF10

2014

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Peptidoglycan (PG) hydrolases associated with bacterial type IV secretion systems (T4SSs) are thought to generate localized lesions in the PG layer to facilitate assembly of the translocation channel. The pheromone-responsive plasmid pCF10 of Enterococcus faecalis encodes a putative cell wall hydrolase, PrgK, and here we report that a prgK deletion abolished functionality of the pCF10-encoded T4SS as monitored by pCF10 conjugative transfer. Expression in trans of wild-type prgK fully complemented this mutation. PrgK has three potential hydrolase motifs resembling staphylococcal LytM, soluble lytic transglycosylase (SLT), and cysteine-, histidine-dependent amidohydrolase/peptidase (CHAP) domains. Complementation analyses with mutant alleles established that PrgK bearing two hydrolase domains in any combination supported near-wild-type plasmid transfer, and PrgK bearing a single hydrolase domain supported at least a low level of transfer in filter matings. When exported to the Escherichia coli periplasm, each domain disrupted cell growth, and combinations of domains additionally induced cell rounding and blebbing and conferred enhanced sensitivity to osmotic shock. Each domain bound PG in vitro, but only the SLT domain exhibited detectable hydrolase activity, as shown by zymographic analyses and release of fluorescent PG fragments. Genes encoding three T4SS-associated, putative hydrolases, Lactococcus lactis CsiA, Tn925 Orf14, and pIP501 TraG, partially complemented the ⌬prgK mutation. Our findings establish that PrgK is an essential component of the pCF10-encoded Prg/Pcf T4SS and that its hydrolase domains coordinate their activities for full PrgK function. PrgK is indispensable for plasmid transfer in liquid matings, suggestive of a role in formation or stabilization of mating junctions.

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“…According to Laverde Gomez et al (27), this gene is located on a megaplasmid, which is widely distributed among clinically associated E. faecium and seems to be restricted to this species.…”

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confidence: 99%

Genotypic Diversity, Antibiotic Resistance and Bacteriocin Production of Enterococci Isolated from Rhizospheres

et al. 2012

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This study aimed to identify and to characterize rhizospheric-derived enterococci. The results showed the prevalence of Enterococcus faecium species (97%) vs. Enterococcus durans (3%). Susceptibility testing for antibiotics showed a low percentage of resistance to erythromycin (3.2%) and tetracycline (11.2%), and intermediate resistance to vancomycin (6.5%). Nevertheless, a high proportion of bacteriocin production was recorded. Furthermore, PCR detection of antibiotic resistance and bacteriocin production-encoding genes was investigated. Pulsed-field gel electrophoresis typing (PFGE) showed a great variability of enterococci in the rhizosphere. Moreover, mutilocus-sequence-typing analysis (MLST) revealed the identification of three new sequence types (STs), which were registered as ST613, ST614 and ST615.

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A multiresistance megaplasmid pLG1 bearing a hylEfm genomic island in hospital Enterococcus faecium isolates

Cited by 71 publications

References 56 publications

Molecular Analysis of VanA Outbreak ofEnterococcus faeciumin Two Warsaw Hospitals: The Importance of Mobile Genetic Elements

Molecular Analysis of VanA Outbreak ofEnterococcus faeciumin Two Warsaw Hospitals: The Importance of Mobile Genetic Elements

PrgK, a Multidomain Peptidoglycan Hydrolase, Is Essential for Conjugative Transfer of the Pheromone-Responsive Plasmid pCF10

Genotypic Diversity, Antibiotic Resistance and Bacteriocin Production of Enterococci Isolated from Rhizospheres

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