Influence of IS
            <i>256</i>
            on Genome Variability and Formation of Small-Colony Variants in Staphylococcus aureus

Kleinert, Franziska; Kallies, René; Hort, Michael; Zweynert, Annegret; Szekat, Christiane; Nagel, Michael; Bierbaum, Gabriele

doi:10.1128/aac.00144-17

Cited by 24 publications

(15 citation statements)

References 61 publications

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“…In all cases, the TSD was 8 bp long and there appeared to be a preference for integration into A/T‐rich sequences. This is consistent with the A/T‐rich 8‐bp TSDs observed in the genome of C. thermocellum and for IS 256 itself (Loessner et al , 2002; Kleinert et al , 2017). Neither TIR junction integration (Appendix Fig S1B) nor Figure‐eight formation (Appendix Fig S1C) was detected using TnpA with the active site mutation D175A.…”

Section: Resultssupporting

confidence: 85%

Structures of IS C th4 transpososomes reveal the role of asymmetry in copy‐out/paste‐in DNA transposition

et al. 2020

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Copy-out/paste-in transposition is a major bacterial DNA mobility pathway. It contributes significantly to the emergence of antibiotic resistance, often by upregulating expression of downstream genes upon integration. Unlike other transposition pathways, it requires both asymmetric and symmetric strand transfer steps. Here, we report the first structural study of a copy-out/paste-in transposase and demonstrate its ability to catalyze all pathway steps in vitro. X-ray structures of ISCth4 transposase, a member of the IS256 family of insertion sequences, bound to DNA substrates corresponding to three sequential steps in the reaction reveal an unusual asymmetric dimeric transpososome. During transposition, an array of N-terminal domains binds a single transposon end while the catalytic domain moves to accommodate the varying substrates. These conformational changes control the path of DNA flanking the transposon end and the generation of DNA-binding sites. Our results explain the asymmetric outcome of the initial strand transfer and show how DNA binding is modulated by the asymmetric transposase to allow the capture of a second transposon end and to integrate a circular intermediate.

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Section: Resultssupporting

confidence: 85%

Structures of IS C th4 transpososomes reveal the role of asymmetry in copy‐out/paste‐in DNA transposition

et al. 2020

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“…2). The IS 256 is an insertion sequence that confers a strong genomic plasticity to MRSA [41]. Since IS 256 is present in multiple copies in the staphylococcal genome [32], the dimension of the polymorphic inter-IS 256 sequences has previously been used as a typing tool for MRSA [42].…”

Section: Discussionmentioning

confidence: 99%

Prevalence, molecular epidemiology, and antimicrobial resistance of methicillin-resistant Staphylococcus aureus from swine in southern Italy

et al. 2019

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Background Colonization by livestock-associated MRSA (LA-MRSA) has increasingly been reported in the swine population worldwide. The aim of this study was to assess the prevalence of MRSA nasal carriage in healthy pigs, including the black ( Calabrese ) breed, from farms in the Calabria Region (Southern Italy). Between January and March 2018, a total of 475 healthy pigs reared in 32 farms were sampled by nasal swabbing. MRSA isolates were characterized by spa , MLST and SCC mec typing, and susceptibility testing to 17 antimicrobials. Results 22 of 32 (66.8%) pig farms resulted positive for MRSA. The prevalence of MRSA was 46.1% (219 MRSA culture-positive out of 475 samples). MRSA colonization was significantly higher in intensive farms and in pigs with a recent or ongoing antimicrobial treatment. All 219 MRSA isolates were assigned to ST398. The most common spa types were t011 (37.0%), t034 (22.4%) and t899 (15.1%). A novel spa type (t18290) was detected in one isolate. An insertion of IS 256 in the ST398-specific A07 fragment of the SAPIG2195 gene was detected in 10 out of 81 t011 isolates. Nearly all isolates carried the SCC mec type V element, except 11 isolates that carried the SCC mec type IVc. None of the isolates was positive for the Panton-Valentine leukocidin. All isolates were resistant to tetracycline. High resistance rates were also found for clindamycin (93.1%), trimethoprim/sulfamethoxazole (68.4%), fluoroquinolones (47.9–65.3%) and erythromycin (46.1%). None of the isolates was resistant to vancomycin and fusidic acid. Overall, a multidrug resistant phenotype was observed in 88.6% of isolates. Conclusions We report a high prevalence of MRSA among healthy swine in Southern Italy farms, with higher isolation frequency associated with intensive farming. The epidemiological types identified in our study reflect those reported in other European countries. Our findings underscore the importance of monitoring the evolution of LA-MRSA in pig farms in order to implement control measures and reduce the risk of spread in the animal population. Electronic supplementary material The online version of this article (10.1186/s12866-019-1422-x) contains supplementary material, which is available to authorized users.

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“…IS256 can shape the genome by affecting gene expression. In several successful virulent MRSA STs, such as ST 247, the presence of IS256 has been linked to increased virulence, vancomycin resistance and formation of small-colony variants (Kleinert et al, 2017). The opposite was demonstrated in Brazilian MRSA ST 239 isolates where IS256 was integrated near global regulatory genes ( agr and mgr ) likely leading to rapid changes of bacterial traits, resulting in reduced virulence (Botelho et al, 2019).…”

Section: Discussionmentioning

confidence: 99%

Comparative Genomic Analysis of Staphylococcus haemolyticus Reveals Key to Hospital Adaptation and Pathogenicity

et al. 2019

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Staphylococcus haemolyticus is a skin commensal gaining increased attention as an emerging pathogen of nosocomial infections. However, knowledge about the transition from a commensal to an invasive lifestyle remains sparse and there is a paucity of studies comparing pathogenicity traits between commensal and clinical isolates. In this study, we used a pan-genomic approach to identify factors important for infection and hospital adaptation by exploring the genomic variability of 123 clinical isolates and 46 commensal S. haemolyticus isolates. Phylogenetic reconstruction grouped the 169 isolates into six clades with a distinct distribution of clinical and commensal isolates in the different clades. Phenotypically, multi-drug antibiotic resistance was detected in 108/123 (88%) of the clinical isolates and 5/46 (11%) of the commensal isolates (p < 0.05). In the clinical isolates, we commonly identified a homolog of the serine-rich repeat glycoproteins sraP. Additionally, three novel capsular polysaccharide operons were detected, with a potential role in S. haemolyticus virulence. Clinical S. haemolyticus isolates showed specific signatures associated with successful hospital adaption. Biofilm forming S. haemolyticus isolates that are resistant to oxacillin (mecA) and aminoglycosides (aacA-aphD) are most likely invasive isolates whereas absence of these traits strongly indicates a commensal isolate. We conclude that our data show a clear segregation of isolates of commensal origin, and specific genetic signatures distinguishing the clinical isolates from the commensal isolates. The widespread use of antimicrobial agents has probably promoted the development of successful hospital adapted clones of S. haemolyticus clones through acquisition of mobile genetic elements or beneficial point mutations and rearrangements in surface associated genes.

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Influence of IS 256 on Genome Variability and Formation of Small-Colony Variants in Staphylococcus aureus

Cited by 24 publications

References 61 publications

Structures of IS C th4 transpososomes reveal the role of asymmetry in copy‐out/paste‐in DNA transposition

Structures of IS C th4 transpososomes reveal the role of asymmetry in copy‐out/paste‐in DNA transposition

Prevalence, molecular epidemiology, and antimicrobial resistance of methicillin-resistant Staphylococcus aureus from swine in southern Italy

Comparative Genomic Analysis of Staphylococcus haemolyticus Reveals Key to Hospital Adaptation and Pathogenicity

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