Cloning of Insertion Sequence IS1485 from Enterococcus Species

Cheng, S; Musso, Richard E.; Li, Rui; Niemeyer, Debra M.; Shaw, Joe J.; McCleskey, F K; DelVecchio, Vito G.

doi:10.1006/plas.1999.1409

Cited by 5 publications

(3 citation statements)

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“…IS 981, which is mostly reported in lactococci, was found in 17/20 E. faecium strains and 12/20 E. faecalis strains. IS 1485, which is widely spread in many enterococcal species [ 95 ], were found in all tested strains. The putative transposase of IS Enfa3 was found in all E. faecium , while it was absent in E. faecalis (Additional file 3 : Figure S2B and Additional file 5 : Figure S3B).…”

Section: Resultsmentioning

confidence: 99%

Investigating the mobilome in clinically important lineages of Enterococcus faecium and Enterococcus faecalis

et al. 2015

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BackgroundThe success of Enterococcus faecium and E. faecalis evolving as multi-resistant nosocomial pathogens is associated with their ability to acquire and share adaptive traits, including antimicrobial resistance genes encoded by mobile genetic elements (MGEs). Here, we investigate this mobilome in successful hospital associated genetic lineages, E. faecium sequence type (ST)17 (n=10) and ST78 (n=10), E. faecalis ST6 (n=10) and ST40 (n=10) by DNA microarray analyses.ResultsThe hybridization patterns of 272 representative targets including plasmid backbones (n=85), transposable elements (n=85), resistance determinants (n=67), prophages (n=29) and clustered regularly interspaced short palindromic repeats (CRISPR)-cas sequences (n=6) separated the strains according to species, and for E. faecalis also according to STs. RCR-, Rep_3-, RepA_N- and Inc18-family plasmids were highly prevalent and with the exception of Rep_3, evenly distributed between the species. There was a considerable difference in the replicon profile, with rep17/pRUM, rep2/pRE25, rep14/EFNP1 and rep20/pLG1 dominating in E. faecium and rep9/pCF10, rep2/pRE25 and rep7 in E. faecalis strains. We observed an overall high correlation between the presence and absence of genes coding for resistance towards antibiotics, metals, biocides and their corresponding MGEs as well as their phenotypic antimicrobial susceptibility pattern. Although most IS families were represented in both E. faecalis and E. faecium, specific IS elements within these families were distributed in only one species. The prevalence of IS256-, IS3-, ISL3-, IS200/IS605-, IS110-, IS982- and IS4-transposases was significantly higher in E. faecium than E. faecalis, and that of IS110-, IS982- and IS1182-transposases in E. faecalis ST6 compared to ST40. Notably, the transposases of IS981, ISEfm1 and IS1678 that have only been reported in few enterococcal isolates were well represented in the E. faecium strains. E. faecalis ST40 strains harboured possible functional CRISPR-Cas systems, and still resistance and prophage sequences were generally well represented.ConclusionsThe targeted MGEs were highly prevalent among the selected STs, underlining their potential importance in the evolution of hospital-adapted lineages of enterococci. Although the propensity of inter-species horizontal gene transfer (HGT) must be emphasized, the considerable species-specificity of these MGEs indicates a separate vertical evolution of MGEs within each species, and for E. faecalis within each ST.Electronic supplementary materialThe online version of this article (doi:10.1186/s12864-015-1407-6) contains supplementary material, which is available to authorized users.

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Section: Resultsmentioning

confidence: 99%

Investigating the mobilome in clinically important lineages of Enterococcus faecium and Enterococcus faecalis

et al. 2015

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“…However, its equivalent in pMG1 additionally contains two ORFs in the central region that correspond to orfA and orfB of IS1485, encoding for transposases. 41 They seem to have been lost through a deletion that turned the element into a mobile insertion cassette where passenger genes with unknown functions are present instead of transposases. 42 This process might be an explanation of how fragment F23-24 ended up in Tn916, turning it into Tn6198.…”

Section: Discussionmentioning

confidence: 99%

Tn6198, a novel transposon containing the trimethoprim resistance gene dfrG embedded into a Tn916 element in Listeria monocytogenes

Bertsch

Uruty

Anderegg

et al. 2013

Journal of Antimicrobial Chemotherapy

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This study reports the first Tn916-like element associated with a trimethoprim resistance gene, as well as the first fully characterized transposon conferring multidrug resistance in L. monocytogenes. This is of concern, as trimethoprim is administered to listeriosis patients with β-lactam allergy and as Tn6198 has a large potential for dissemination, indicated by both intra-species and inter-genus transfer.

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“…The DNA sequence of the ORF5–ORF6 region (3921–5286 bp of the pMG1 map) was identical to IS 1485 , which was originally identified in E. faecium , and ORF5 and ORF6 corresponded to orfA and orfB of IS 1485 , respectively (Cheng et al , 1999). IS 1485 was flanked by a 4‐bp direct repeat (TATC) at both ends.…”

Section: Resultsmentioning

confidence: 85%

Complete nucleotide sequencing and analysis of the 65-kb highly conjugativeEnterococcus faeciumplasmid pMG1: identification of the transfer-related region and the minimum region required for replication

Tanimoto

Ike

2008

FEMS Microbiology Letters

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pMG1 (65.1 kb) is a highly conjugative, pheromone-independent Enterococcus faecium plasmid that carries a Tn4001-like transposon encoding gentamicin resistance. The complete nucleotide sequence (65 029 bp) of the pMG1 plasmid was determined and 73 ORFs lying in the same transcription orientation were identified. Sixty-one of the 73 ORFs showed a high degree of similarity (90-100% identity at the amino acid level) to the ORFs of the pHTbeta plasmids. Like the pHTbeta plasmid, 22 of the pMG1 plasmid ORFs showed homology with ORFs present on the pXO2 plasmid (96.2 kb), which is the virulence plasmid essential for capsular formation by Bacillus anthracis. Analysis of tra mutants created by Tn917 insertion and Northern analysis of transcripts indicated that ORFs 15-49, lying in the 31.7 kb region between 13.6 and 45.3 kb on the plasmid map, were related to transfer. This region was designated as the Tra I region of pMG1. A 5.9-kb HindIII fragment that replicates autonomously in Enterococcus faecalis was cloned and analysis of this fragment by deletion and in vitro insertion mutations showed that ORF10 (rep) and the inverted repeat sequence in the noncoding region between ORF8 and ORF9 were necessary for pMG1 replication.

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Cloning of Insertion Sequence IS1485 from Enterococcus Species

Cited by 5 publications

References 4 publications

Investigating the mobilome in clinically important lineages of Enterococcus faecium and Enterococcus faecalis

Investigating the mobilome in clinically important lineages of Enterococcus faecium and Enterococcus faecalis

Tn6198, a novel transposon containing the trimethoprim resistance gene dfrG embedded into a Tn916 element in Listeria monocytogenes

Complete nucleotide sequencing and analysis of the 65-kb highly conjugativeEnterococcus faeciumplasmid pMG1: identification of the transfer-related region and the minimum region required for replication

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