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1.
Plasmids carrying two IS30 elements in the same orientation, as in the composite transposon Tn2706, are structurally unstable in Escherichia coli. A primary segregation product is formed by site-specific deletion of the sequences carried between the two IS30 elements. The resulting covalently closed replicon carries the two IS30 elements as tandem repeats separated by only 2 bp. This (IS30)2 structure is extremely unstable, but it can nevertheless be isolated on its vector plasmid and, after purification, can be reintroduced into host cells by transformation. Among the descendants of transformants of recA
– bacteria, replicated copies of the introduced (IS30)2 structure are still present, together with various kinds of segregation products which provide evidence for the efficient generation of DNA rearrangements. Most abundant is the product of another site-specific recombination between two identical ends of the IS30 elements involved, which results in the presence of just one intact IS30 on the plasmid. Apart from this, and depending on the presence of appropriate targets for IS30 transposition, various transposition products of (IS30)2 are also seen. Intramolecular reactions lead to DNA inversions and deletions with breakpoints other than IS30 ends. In intermolecular reactions inverse transposition occurs at high frequency and one also obtains simple transposition and cointegration. A mutational study revealed the requirement in cis of one intact IS30 transposase gene and of both proximal ends of the two IS30 elements concerned not only for the formation of (IS30)2, but also for its further rearrangement reactions, including the efficient formation of site-specific deletions. A model is proposed, which postulates that (IS30)2 intermediates play a key role in IS30 transposition pathways in which the formation of (IS30)2 may be rate-limiting. Once this structure is formed, it gives rise to a burst of transpositional rearrangements in the subclone carrying (IS30)2. Evolutionary implications of these findings are discussed. 相似文献
2.
IIdikó Szeverényi Tamás Bodoky Ferenc Olasz 《Molecular genetics and genomics : MGG》1996,251(3):281-289
We have isolated and characterized a dimer derivative of the extensively studiedEscherichia coli insertion sequence IS2. The dimer structure — called (IS2)2 — consists of two IS2 elements arranged as a direct repeat, separated by 1 bp. The junction between the (IS2)2 dimer and target sequences is located at various positions in independent isolates; however, one position was preferred. The transposition of (IS2)2 into a target plasmid resulted in cointegrate-type structures. The transposition frequency of the (IS2)2 dimer itself was significantly higher than that of the isogenic monomer IS2 insertion. The poor stability and high activity of (IS2)2 indicates that this is an active transposition intermediate. The mode of transposition of (IS2)2 is analogous to the joined dimer model described in the case of (IS21)2 and (IS30)2. 相似文献
3.
The mobile element IS30 has 26-bp imperfect terminal inverted repeats (IRs) that are indispensable for transposition. We have analyzed the effects of IR mutations on both major transposition steps, the circle formation and integration of the abutted ends, characteristic for IS30. Several mutants show strikingly different phenotypes if the mutations are present at one or both ends and differentially influence the transposition steps. The two IRs are equivalent in the recombination reactions and contain several functional regions. We have determined that positions 20 to 26 are responsible for binding of the N-terminal domain of the transposase and the formation of a correct 2-bp spacer between the abutted ends. However, integration is efficient without this region, suggesting that a second binding site for the transposase may exist, possibly within the region from 4 to 11 bp. Several mutations at this part of the IRs, which are highly conserved in the IS30 family, considerably affected both major transposition steps. In addition, positions 16 and 17 seem to be responsible for distinguishing the IRs of related insertion sequences by providing specificity for the transposase to recognize its cognate ends. Finally, we show both in vivo and in vitro that position 3 has a determining role in the donor function of the ends, especially in DNA cleavage adjacent to the IRs. Taken together, the present work provides evidence for a more complex organization of the IS30 IRs than was previously suggested.Mobile DNA elements have been described in most organisms and represent a considerable proportion of their genetic material. These elements play an important role in the evolution of the host genome due to their capacities to generate DNA rearrangements and influence the expression of neighboring genes. Their ability to form compound transposons contributes to the sequestering and dispersion of accessory genes, such as those specifying resistance to antibiotics, virulence, and various catabolic activities. The simplest mobile elements are the bacterial insertion sequences (ISs), which typically harbor one or two open reading frames (ORF) coding for the transposase (Tpase). More than 2,400 ISs have been described and classified into families (IS Finder, http://www-is.biotoul.fr/) on the basis of similarities in their genetic organization and Tpases (30). The terminal inverted repeats (IRs) are essential for the transposition of most ISs. The IRs, together with the Tpase, form a complex where the cleavage and strand transfer reactions occur. The IRs generally contain two functional modules: the internal region serves as the binding site of Tpase, while the terminal part is required for DNA cleavage and the strand transfer process (2). Besides these principal cis-acting elements, some ISs carry additional regulatory DNA sequences in the IRs or in the subterminal regions (18).The IS30 family currently comprises more than 80 elements distributed throughout the Gram-positive and Gram-negative bacteria and the Archaea (IS Finder, http://www-is.biotoul.fr). IS30 (1, 5), the founding element of the family, is 1,221 bp long and has 26-bp imperfect IRs (the left end of the IR [IRL] and the right end of the IR [IRR]; Fig. Fig.1A)1A) and one ORF with a coding capacity for a 44.3-kDa Tpase. The element has a preference for two distinct types of target sequences: the natural hot spots (HSs), characterized by a 24-bp symmetric consensus (23), and the IRs of the element itself (21, 22). Potential helix-turn-helix motifs (HTH) responsible for HS and IR targeting are located in the N-terminal region of the Tpase (19). While the first motif, HTH1, is required only for transposition into the HS sequences, the conserved H-HTH2 motif is essential for both IR and HS targeting (15, 19).Open in a separate windowFIG. 1.Transposition assays for comparing the IS30-based transposons composed of simple IRs. (A) Comparison of the IS30 IR sequences. Dots indicate matching bases. (B) Schematic representation of the intermolecular transposition pathway. The graph shows the two major steps characteristic for IS30 transposition (steps 1 and 2). The transposon donor plasmid and its derivative, the circular transposon (thin line), carry the 26-bp IRs of IS30 (boxes with open and filled triangles representing IRL and IRR, respectively). The Cmr gene flanking the transposon in the donor plasmid is shown as a gray box. The target plasmid (dotted line) carries the GOHS hot spot sequence (cross-hatched box). (C) Transposition frequencies of IS30-based transposons with different combinations of the IRs. The graph shows the overall frequency of transposition into the hot spot (steps 1 and 2) and the frequency of the major steps assayed separately. Data were obtained from at least three parallel experiments.IS30 transposition occurs through two major steps (14, 24) (Fig. (Fig.1B).1B). The first is the formation of an active intermediate by joining of the IRs. This process involves the Tpase-catalyzed cleavage of one strand at the 3′ IS end, which then attacks the same strand 2 bp outside the other IR. This strand transfer generates a single-strand bridge between the ends and leads to a figure-eight structure (33). This active transposition intermediate carrying the joined IRs probably proceeds via replicative resolution, as described for IS911 (11, 25) and IS2 (16). The resolution can lead to the circularization of a single IS or to the formation of a head-to-tail repeat of two IS30 copies. In the second step of transposition, the active forms interact with the target DNA, resulting in the known transposition products: simple insertion, deletion, inversion, or replicon fusion (14, 24).In this work, we describe the modularity of the IR ends of IS30 by analyzing several mutants. According to our results, the IS30 IRs can be divided into functional regions that are differently involved in the main transposition steps. We show that positions 2 and 3 play a pivotal role in cleavage of the ends and, consequently, in their donor function. While the terminal part (1 to 17 bp) of the IRs is indispensable for both major steps, the internal region, i.e., the binding site for the N-terminal part of Tpase (20 to 26 bp), appears to be required only for the junction formation. Although the exact role of the terminal part of IRs is less clear, several mutations in this region considerably affected both the junction formation and integration. The fact that the internal IR region is not involved in the integration suggests that the Tpase binds to other sequences during this reaction. 相似文献
4.
Inverted repeats of insertion sequences (ISs) are indispensable for transposition. We demonstrate that sub-terminal sequences adjacent to the inverted repeats of IS30 are also required for optimal transposition activity. We have developed a cell-free recombination system and showed that the transposase catalyses formation of a figure-of-eight transposition intermediate, where a 2 bp long single strand bridge holds the inverted repeat sequences (IRs) together. This is the first demonstration of the figure-of-eight structure in a non-IS3 family element, suggesting that this mechanism is likely more widely adopted among IS families. We show that the absence of sub-terminal IS30 sequences negatively influences figure-of-eight production both in vivo and in vitro. These regions enhance IR-IR junction formation and IR-targeting events in vivo. Enhancer elements have been identified within 51 bp internal to IRL and 17 bp internal to IRR. In the right end, a decanucleotide, 5′-GAGATAATTG-3′, is responsible for wild-type activity, while in the left end, a complex assembly of repetitive elements is required. Functioning of the 10 bp element in the right end is position-dependent and the repetitive elements in the left end act cooperatively and may influence bendability of the end. In vitro kinetic experiments suggest that the sub-terminal enhancers may, at least partly, be transposase-dependent. Such enhancers may reflect a subtle regulatory mechanism for IS30 transposition. 相似文献
5.
Macrotransposition and other complex chromosomal restructuring in maize by closely linked transposons in direct orientation 总被引:2,自引:1,他引:1
Several observations indicate that compatible ends of separate, yet closely linked, transposable elements (TEs) can interact in alternative transposition reactions. First, pairs of TEs cause chromosome breaks with frequencies inversely related to the intertransposon distance. Second, some combinations of two TEs produce complex rearrangements that often include DNA adjacent to one or both elements. In pairs of TEs in direct orientation, alternative reactions involving the external ends of the two TEs should lead to the transposition of a macrotransposon consisting of both elements plus the intervening chromosomal segment. Such macrotransposons have been hypothesized previously based on deletions, but no macrotransposon insertions have been recovered. To detect macrotransposition, we have analyzed heritable chromosomal rearrangements produced by a chromosome-breaking pair of Ac and Ds elements situated 6.5 kb apart in direct orientation in a part of the maize (Zea mays) genome dispensable for viability. Here, we show that the postulated macrotransposon can excise and reinsert elsewhere in the genome. In addition, this transposon pair produces other complex rearrangements, including deletions, inversions, and reshuffling of the intertransposon segment. Thus, closely linked TE pairs, a common transposition outcome in some superfamilies, are adept at restructuring chromosomes and may have been instrumental in reshaping plant genomes. 相似文献
6.
The maize Activator/Dissociation (Ac/Ds) elements are members of the hAT (hobo, Ac, and Tam3) superfamily of type II (DNA) transposons that transpose through a “cut-and-paste” mechanism. Previously, we reported that a pair of Ac ends in reversed orientation is capable of undergoing alternative transposition reactions that can generate large-scale chromosomal rearrangements, including deletions and inversions. We show here that rearrangements induced by reversed Ac ends transposition can join the coding and regulatory sequences of two linked paralogous genes to generate a series of chimeric genes, some of which are functional. To our knowledge, this is the first report demonstrating that alternative transposition reactions can recombine gene segments, leading to the creation of new genes. 相似文献
7.
We have isolated and characterized a dimer derivative of the extensively studiedEscherichia coli insertion sequence IS2. The dimer structure — called (IS2)2 — consists of two IS2 elements arranged as a direct repeat, separated by 1 bp. The junction between the (IS2)2 dimer and target sequences is located at various positions in independent isolates; however, one position was preferred. The transposition of (IS2)2 into a target plasmid resulted in cointegrate-type structures. The transposition frequency of the (IS2)2 dimer itself was significantly higher than that of the isogenic monomer IS2 insertion. The poor stability and high activity of (IS2)2 indicates that this is an active transposition intermediate. The mode of transposition of (IS2)2 is analogous to the joined dimer model described in the case of (IS21)2 and (IS30)2. 相似文献
8.
IS6110-Mediated Deletions of Wild-Type Chromosomes of Mycobacterium tuberculosis 总被引:1,自引:0,他引:1
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Z. Fang C. Doig D. T. Kenna N. Smittipat P. Palittapongarnpim B. Watt K. J. Forbes 《Journal of bacteriology》1999,181(3):1014-1020
The ipl locus is a site for the preferential insertion of IS6110 and has been identified as an insertion sequence, IS1547, in its own right. Various deletions around the ipl locus of clinical isolates of Mycobacterium tuberculosis were identified, and these deletions ranged in length from several hundred base pairs up to several kilobase pairs. The most obvious feature shared by these deletions was the presence of an IS6110 copy at the deletion sites, which suggested two possible mechanisms for their occurrence, IS6110 transposition and homologous recombination. To clarify the mechanism, an investigation was conducted; the results suggest that although deletion transpositionally mediated by IS6110 was a possibility, homologous recombination was a more likely one. The implications of such chromosomal rearrangements for the evolution of M. tuberculosis, for IS6110-mediated mutagenesis, and for the development of genetic tools are discussed. The deletion of genomic DNA in isolates of M. tuberculosis has previously been noted at only a few sites. This study examined the deletional loss of genetic material at a new site and suggests that such losses may occur elsewhere too and may be more prevalent than was previously thought. Distinct from the study of laboratory-induced mutations, the detailed analysis of clinical isolates, in combination with knowledge of their evolutionary relationships to each other, gives us the opportunity to study mutational diversity in isolates that have survived in the human host and therefore offers a different perspective on the importance of particular genetic markers in pathogenesis. 相似文献
9.
The relative integrative recombination frequencies of the F plasmid IS2 and IS3 elements were determined at two Escherichia coli K-12 chromosomal sites by hybridization analysis of Hfr DNA. The sequence organizations of the independent Hfr strains formed by F integration atα3β3 indicate that the relative recombinational frequencies at the two F plasmid IS3 elementsα1β1 and α2β2 are not significantly different. A comparison of the relative recombinational frequencies of the IS2 and IS3 elements of F was provided by analysis of DNA from Hfr strains having F integrated betweenlacandproC (i.e., at the IS2 or at the IS3 element (α5β5) located in this region). No instances of F integration atα5β5 were detected, indicating that integrative recombination at IS2 is significantly more frequent than at IS3 in this chromosomal region. 相似文献
10.
The Clostridium perfringens enterotoxin gene is on a transposon-like element, Tn5565, integrated in the chromosome in human food poisoning strains. The flanking IS elements, IS1470 A and B, are related to IS30. The IS element found in the transposon, IS1469, is related to IS200 and has been found upstream of cpe in all Type A strains. PCR and sequencing studies from cell extracts and plasmid isolations of C. perfringens indicate that Tn5565 can form a circular form with the tandem repeat (IS1470)2, similar to the transposition intermediates described for a number of IS elements. 相似文献
11.
Two Amino Acid Residues of Transposase Contributing to Differential Transposability of IS1 Elements in Escherichia coli
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Escherichia coli W3110 contains four types of IS1 elements in the chromosome. Using an insertion element entrapping system, we collected 116 IS1 plasmid insertion mutants, which resulted from a minimum of 26 independent IS1 insertion events. All of them had insertions of IS1 of the IS1A (IS1E and IS1G) type. Inspection of the transposase sequences of the four IS1 types and the IS1 of the resistance plasmid R100 showed that two amino acid residues, His-193 and Leu-217 of transposase, might contribute to differential transposability of IS1 elements in W3110. The two amino acid residues of the transposase in IS1A (IS1E and IS1G) were altered separately by site-directed mutagenesis, and each mutant was found to mediate transposition at a frequency about 30-fold lower than that of IS1A (IS1E and IS1G). Thus, the assumption that His-193 and Leu-217 of transposase contribute to differential transposability of IS1 elements in W3110 was confirmed. 相似文献
12.
René Rezsöhazy Bernard Hallet Jean Delcour Jacques Mahillon 《Molecular microbiology》1993,9(6):1283-1295
The eight IS 231 variants characterized so far (IS 231 A-F, V and W) display similar transposases with an overall 40% identity. Comparison with all the proka-ryotic transposable elements sequenced so far revealed that the IS231 transposases share two conserved regions with those of 35 other insertion sequences of wide origins. These insertion sequences, defining the IS4 family, have a common bipartite organization of their ends and are divided into two similarity groups. Interestingly, the transposase domains conserved within this family display similarities with the well known integrase domain shared by transposases of the IS3 and IS15 families, and integrases of retroelements. This domain is also found in IS30- related elements and Tn7 TnsB protein. Amino acid residues conserved throughout all these prokaryotic and eukaryotic mobile genetic elements define a major transposase/integrase motif, likely to play an important role in the transposition process. 相似文献
13.
In the present study we report on the excision of IS30 elements and IS30-derived composite transposons. Frequent loss of IS30 was observed during dissolution of dimeric IS30 structures, containing IR–IR junctions, leading to resealed donor molecules. In contrast, unambiguous transpositional excision resulting in resealed remainder products could not be identified in the case of a monomeric element. The bias in the excision of monomeric and dimeric IS30 structures indicates a difference in the molecular mechanism of transposition of IS30 monomers and dimers. Sequence data on the rarely detected plasmids missing full IS or Tn copies rather suggest that all products were derived from illegitimate recombination. The reaction occurred between short homologies and was independent of the transposase activity. Similar IS30 excision events accompanied by multiple plasmid or genome rearrangements were detected in Pseudomonas putida and Rhizobium meliloti, yielding stable replicons that retained the selective marker gene of the transposon. We provide evidence that both transposition and illegitimate recombination can contribute to the stabilization of replicons through the elimination of IS elements, which emphasizes the evolutionary significance of these events. 相似文献
14.
IS117, the 2.6 kb mini-circle of Streptomyces coelicolor A3(2), is a transposable element previously shown to be integrated into two distant sites in the chromosome. When introduced into S. lividans, IS117 integrates into one preferred chromosomal site, but when this site was artificially deleted, IS117 integrated into many secondary sites. Nucleotide sequence analysis of several secondary integration sites revealed varying degrees of similarity with the preferred site, but no consensus sequence. Nevertheless, sites more similar to the preferred site tended to be occupied more often than those that are less similar. Insertion of IS117 into secondary sites in the chromosome of S. lividans sometimes mediated chromosomal rearrangements. It was shown that some strains containing IS117 integrated into secondary sites had suffered deletions of chromosomal DNA. Deletions were adjacent to the inserted element and were at least several kilobases long. The proposed model implicates homologous recombination between IS117 copies integrated into two different secondary sites in the same chromosome as a cause of the deletions. 相似文献
15.
ISLpl1 Is a Functional IS30-Related Insertion Element in Lactobacillus plantarum That Is Also Found in Other Lactic Acid Bacteria
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We describe the first functional insertion sequence (IS) element in Lactobacillus plantarum. ISLpl1, an IS30-related element, was found on the pLp3 plasmid in strain FB335. By selection of spontaneous mutants able to grow in the presence of uracil, it was demonstrated that the IS had transposed into the uracil phosphoribosyltransferase-encoding gene upp on the FB335 chromosome. The plasmid-carried IS element was also sequenced, and a second potential IS element was found: ISLpl2, an IS150-related element adjacent to ISLpl1. When Southern hybridization was used, the copy number and genome (plasmid versus chromosome) distribution data revealed different numbers and patterns of ISLpl1-related sequences in different L. plantarum strains as well as in Pediococcus strains. The ISLpl1 pattern changed over many generations of the strain L. plantarum NCIMB 1406. This finding strongly supports our hypothesis that ISLpl1 is a mobile element in L. plantarum. Database analysis revealed five quasi-identical ISLpl1 elements in Lactobacillus, Pediococcus, and Oenococcus strains. Three of these elements may be cryptic IS, since point mutations or 1-nucleotide deletions were found in their transposase-encoding genes. In some cases, ISLpl1 was linked to genes involved in cold shock adaptation, bacteriocin production, sugar utilization, or antibiotic resistance. ISLpl1 is transferred among lactic acid bacteria (LAB) and may play a role in LAB genome plasticity and adaptation to their environment. 相似文献
16.
IS911 is a bacterial insertion sequence composed of two consecutive overlapping open reading frames (ORFs [orfA and orfB]) encoding the transposase (OrfAB) as well as a regulatory protein (OrfA). These ORFs are bordered by terminal left and right inverted repeats (IRL and IRR, respectively) with several differences in nucleotide sequence. IS911 transposition is asymmetric: each end is cleaved on one strand to generate a free 3′-OH, which is then used as the nucleophile in attacking the opposite insertion sequence (IS) end to generate a free IS circle. This will be inserted into a new target site. We show here that the ends exhibit functional differences which, in vivo, may favor the use of one compared to the other during transposition. Electromobility shift assays showed that a truncated form of the transposase [OrfAB(1-149)] exhibits higher affinity for IRR than for IRL. While there was no detectable difference in IR activities during the early steps of transposition, IRR was more efficient during the final insertion steps. We show here that the differential activities between the two IRs correlate with the different affinities of OrfAB(1-149) for the IRs during assembly of the nucleoprotein complexes leading to transposition. We conclude that the two inverted repeats are not equivalent during IS911 transposition and that this asymmetry may intervene to determine the ordered assembly of the different protein-DNA complexes involved in the reaction. 相似文献
17.
18.
Heewook Lee Thomas G. Doak Ellen Popodi Patricia L. Foster Haixu Tang 《Nucleic acids research》2016,44(15):7109-7119
A majority of large-scale bacterial genome rearrangements involve mobile genetic elements such as insertion sequence (IS) elements. Here we report novel insertions and excisions of IS elements and recombination between homologous IS elements identified in a large collection of Escherichia coli mutation accumulation lines by analysis of whole genome shotgun sequencing data. Based on 857 identified events (758 IS insertions, 98 recombinations and 1 excision), we estimate that the rate of IS insertion is 3.5 × 10−4 insertions per genome per generation and the rate of IS homologous recombination is 4.5 × 10−5 recombinations per genome per generation. These events are mostly contributed by the IS elements IS1, IS2, IS5 and IS186. Spatial analysis of new insertions suggest that transposition is biased to proximal insertions, and the length spectrum of IS-caused deletions is largely explained by local hopping. For any of the ISs studied there is no region of the circular genome that is favored or disfavored for new insertions but there are notable hotspots for deletions. Some elements have preferences for non-coding sequence or for the beginning and end of coding regions, largely explained by target site motifs. Interestingly, transposition and deletion rates remain constant across the wild-type and 12 mutant E. coli lines, each deficient in a distinct DNA repair pathway. Finally, we characterized the target sites of four IS families, confirming previous results and characterizing a highly specific pattern at IS186 target-sites, 5′-GGGG(N6/N7)CCCC-3′. We also detected 48 long deletions not involving IS elements. 相似文献
19.
Geisa A. L. Gonçalves Pedro H. Oliveira Ana G. Gomes Kristala L. J. Prather Leslie A. Lewis Duarte M. F. Prazeres Gabriel A. Monteiro 《Applied microbiology and biotechnology》2014,98(15):6609-6619
Insertion specificity of mobile genetic elements is a rather complex aspect of DNA transposition, which, despite much progress towards its elucidation, still remains incompletely understood. We report here the results of a meta-analysis of IS2 target sites from genomic, phage, and plasmid DNA and find that newly acquired IS2 elements are consistently inserted around abrupt DNA compositional shifts, particularly in the form of switch sites of GC skew. The results presented in this study not only corroborate our previous observations that both the insertion sequence (IS) minicircle junction and target region adopt intrinsically bent conformations in IS2, but most interestingly, extend this requirement to other families of IS elements. Using this information, we were able to pinpoint regions with high propensity for transposition and to predict and detect, de novo, a novel IS2 insertion event in the 3′ region of the gfp gene of a reporter plasmid. We also found that during amplification of this plasmid, process parameters such as scale, culture growth phase, and medium composition exacerbate IS2 transposition, leading to contamination levels with potentially detrimental clinical effects. Overall, our findings provide new insights into the role of target DNA structure in the mechanism of transposition of IS elements and extend our understanding of how culture conditions are a relevant factor in the induction of genetic instability. 相似文献
20.
Wei-Chun Hung Tomomi Takano Wataru Higuchi Yasuhisa Iwao Olga Khokhlova Lee-Jene Teng Tatsuo Yamamoto 《PloS one》2012,7(10)
Methicillin-resistant Staphylococcus aureus (MRSA) with ST59/SCCmecV and Panton-Valentine leukocidin gene is a major community-acquired MRSA (CA-MRSA) lineage in Taiwan and has been multidrug-resistant since its initial isolation. In this study, we studied the acquisition mechanism of multidrug resistance in an ST59 CA-MRSA strain (PM1) by comparative genomics. PM1’s non-β-lactam resistance was encoded by two unique genetic traits. One was a 21,832-bp composite mobile element structure (MESPM1), which was flanked by direct repeats of enterococcal IS1216V and was inserted into the chromosomal sasK gene; the target sequence (att) was 8 bp long and was duplicated at both ends of MESPM1. MESPM1 consisted of two regions: the 5′-end side 12.4-kb region carrying Tn551 (with ermB) and Tn5405-like (with aph[3′]-IIIa and aadE), similar to an Enterococcus faecalis plasmid, and the 3′-end side 6,587-bp region (MEScat) that carries cat and is flanked by inverted repeats of IS1216V. MEScat possessed att duplication at both ends and additional two copies of IS1216V inside. MESPM1 represents the first enterococcal IS1216V-mediated composite transposon emerged in MRSA. IS1216V-mediated deletion likely occurred in IS1216V-rich MESPM1, resulting in distinct resistance patterns in PM1-derivative strains. Another structure was a 6,025-bp tet-carrying element (MEStet) on a 25,961-bp novel mosaic penicillinase plasmid (pPM1); MEStet was flanked by direct repeats of IS431, but with no target sequence repeats. Moreover, the PM1 genome was deficient in a copy of the restriction and modification genes (hsdM and hsdS), which might have contributed to the acquisition of enterococcal multidrug resistance. 相似文献