The ISApl12 Dimer Circular Intermediate Participates in mcr-1 Transposition

He, Yu-Zhang; Li, Xingping; Miao, Yuanyuan; Lin, Jun; Sun, Ruan-Yang; Wang, Xiao-Pei; Guo, Yaya; Liao, Xiao‐Ping; Liu, Yahong; Feng, Youjun; Sun, Jian

doi:10.3389/fmicb.2019.00015

Cited by 31 publications

(26 citation statements)

References 29 publications

(46 reference statements)

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“…To confirm the IS CR2 -mediated tet (X) transfer, a transposition assay of tet (X) was conducted as previously reported for mcr-1 [ 32 ]. The putative transposition unit of tet (X3) in Clade_U6 10FS3-1, namely Δ tpnF - tet (X3)-hp-hp-IS CR2 , was cloned in a suicide plasmid pSV03 using the ClonExpress Ultra One Step Cloning Kit (Vazyme, China) (Additional file 1 : Table S1) and then transformed into competent E. coli WM3064.…”

Section: Methodsmentioning

confidence: 99%

Genetic diversity and characteristics of high-level tigecycline resistance Tet(X) in Acinetobacter species

Chen

Cui

et al. 2020

Genome Med

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Background The recent emergence and dissemination of high-level mobile tigecycline resistance Tet(X) challenge the clinical effectiveness of tigecycline, one of the last-resort therapeutic options for complicated infections caused by multidrug-resistant Gram-negative and Gram-positive pathogens. Although tet(X) has been found in various bacterial species, less is known about phylogeographic distribution and phenotypic variance of different genetic variants. Methods Herein, we conducted a multiregional whole-genome sequencing study of tet(X)-positive Acinetobacter isolates from human, animal, and their surrounding environmental sources in China. The molecular and enzymatic features of tet(X) variants were characterized by clonal expression, microbial degradation, reverse transcription, and gene transfer experiments, while the tet(X) genetic diversity and molecular evolution were explored by comparative genomic and Bayesian evolutionary analyses. Results We identified 193 tet(X)-positive isolates from 3846 samples, with the prevalence ranging from 2.3 to 25.3% in nine provinces in China. The tet(X) was broadly distributed in 12 Acinetobacter species, including six novel species firstly described here. Besides tet(X3) (n = 188) and tet(X4) (n = 5), two tet(X5) variants, tet(X5.2) (n = 36) and tet(X5.3) (n = 4), were also found together with tet(X3) or tet(X4) but without additive effects on tetracyclines. These tet(X)-positive Acinetobacter spp. isolates exhibited 100% resistance rates to tigecycline and tetracycline, as well as high minimum inhibitory concentrations to eravacycline (2–8 μg/mL) and omadacycline (8–16 μg/mL). Genetic analysis revealed that different tet(X) variants shared an analogous ISCR2-mediated transposon structure. The molecular evolutionary analysis indicated that Tet(X) variants likely shared the same common ancestor with the chromosomal monooxygenases that are found in environmental Flavobacteriaceae bacteria, but sequence divergence suggested separation ~ 9900 years ago (7887 BC), presumably associated with the mobilization of tet(X)-like genes through horizontal transfer. Conclusions Four tet(X) variants were identified in this study, and they were widely distributed in multiple Acinetobacter spp. strains from various ecological niches across China. Our research also highlighted the crucial role of ISCR2 in mobilizing tet(X)-like genes between different Acinetobacter species and explored the evolutionary history of Tet(X)-like monooxygenases. Further studies are needed to evaluate the clinical impact of these mobile tigecycline resistance genes.

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Section: Methodsmentioning

confidence: 99%

Genetic diversity and characteristics of high-level tigecycline resistance Tet(X) in Acinetobacter species

Chen

Cui

et al. 2020

Genome Med

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“…Some studies illustrate that interactions between physiological characteristics and the intestinal metagenome function of an individual are attributed to bacterial community diversity or some specific bacterial groups, e.g., species or functional clusters [ 28 , 29 ]. Previous studies have suggested associations between intestinal metagenome and environmental fluctuation, for example, the gut microbial community changes with habitat fragmentation and degradation [ 30 , 31 , 32 ], altitude [ 33 , 34 ], season [ 35 ], and climate change [ 36 , 37 ] of locations where the hosts live. Cold acclimated laboratory mice experience a dramatic change in gut microbiota composition as temperature increases [ 21 ].…”

Section: Introductionmentioning

confidence: 99%

Habitat Elevation Shapes Microbial Community Composition and Alter the Metabolic Functions in Wild Sable (Martes zibellina) Guts

Liu

Jin

et al. 2021

Animals

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In recent decades, wild sable (Carnivora Mustelidae Martes zibellina) habitats, which are often natural forests, have been squeezed by anthropogenic disturbances such as clear-cutting, tilling and grazing. Sables tend to live in sloped areas with relatively harsh conditions. Here, we determine effects of environmental factors on wild sable gut microbial communities between high and low altitude habitats using Illumina Miseq sequencing of bacterial 16S rRNA genes. Our results showed that despite wild sable gut microbial community diversity being resilient to many environmental factors, community composition was sensitive to altitude. Wild sable gut microbial communities were dominated by Firmicutes (relative abundance 38.23%), followed by Actinobacteria (30.29%), and Proteobacteria (28.15%). Altitude was negatively correlated with the abundance of Firmicutes, suggesting sable likely consume more vegetarian food in lower habitats where plant diversity, temperature and vegetation coverage were greater. In addition, our functional genes prediction and qPCR results demonstrated that energy/fat processing microorganisms and functional genes are enriched with increasing altitude, which likely enhanced metabolic functions and supported wild sables to survive in elevated habitats. Overall, our results improve the knowledge of the ecological impact of habitat change, providing insights into wild animal protection at the mountain area with hash climate conditions.

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“…In brief, we synthesized a cassette in which the mcr-2 and orf genes are flanked by IS Ec69 on both sides and cloned into the vector pUC57, generating the plasmid pUC57- mcr-2 . R6K- Bam HI and R6K- Eco RI primers ( Table 2 ) were used to PCR amplify the backbone using pSV03 ( He et al, 2019 ) as a template which contained the conditional replication origin R6K and Cm R (chloramphenicol) resistance. The amplicon was sub-cloned into the Bam HI and Eco RI restriction sites in pUC57 that contained Tn 7052 to obtain recombinant plasmid pJS05 ( Supplementary Figure 1A ).…”

Section: Methodsmentioning

confidence: 99%

“…The transfer of antimicrobial resistance genes has been found to be catalyzed by specific mobile elements such as insertion element and integron. For instance, IS Apl1 , originally found in Actinobacillus pleuropneumoniae , is located upstream of the mcr-1 gene in the first described mcr-1 -harboring IncI2 type plasmid pHNSHP45 ( Liu et al, 2015 ; Ye et al, 2016 ) and is able to mediate mcr-1 mobilization ( Poirel et al, 2017 ; He et al, 2019 ). Similarly, IS Ec69 , encoding a 217-amino-acid-long DDE-type transposase, belongs to the IS 1595 family and is flanked by two inverted repeats, which share 16/19 nucleotide identity to inverted repeats from the IS 1016C2 transposase.…”

Section: Introductionmentioning

confidence: 99%

ISEc69-Mediated Mobilization of the Colistin Resistance Gene mcr-2 in Escherichia coli

Long

et al. 2021

Front. Microbiol.

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ObjectivesThe emergence of mobile colistin resistance genes has compromised the efficacy of the last resort antibiotic, colistin, in clinical treatment. The mcr-2 gene was first identified in Belgium in association with the insertion sequence ISEc69. However, the molecular mechanisms of mcr-2 mobilization are not well understood.MethodsTo further explore the mobilization of mcr-2 gene via ISEc69, we constructed a conjugative plasmid that carries an intact composite transposon Tn7052. Transposition assays were performed by conjugation, the transposition sites were characterized by arbitrary primed PCR and DNA sequencing.ResultsIn this study, we experimentally demonstrated that mcr-2 could be mobilized as a composite transposon Tn7052 and its transposition generated 8-bp AT-rich duplications in the host genome.ConclusionThese results indicate that mcr-2 gene could be mobilized by ISEc69, the current investigations provide mechanistic insights in the transposition of mcr-2.

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The ISApl12 Dimer Circular Intermediate Participates in mcr-1 Transposition

Cited by 31 publications

References 29 publications

Genetic diversity and characteristics of high-level tigecycline resistance Tet(X) in Acinetobacter species

Genetic diversity and characteristics of high-level tigecycline resistance Tet(X) in Acinetobacter species

Habitat Elevation Shapes Microbial Community Composition and Alter the Metabolic Functions in Wild Sable (Martes zibellina) Guts

ISEc69-Mediated Mobilization of the Colistin Resistance Gene mcr-2 in Escherichia coli

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