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Insertion sequences are compact and pervasive transposable elements found in bacteria, which encode only the genes necessary for their mobilization and maintenance. IS200- and IS605-family transposons undergo 'peel-and-paste' transposition catalysed by a TnpA transposase, but they also encode diverse, TnpB- and IscB-family proteins that are evolutionarily related to the CRISPR-associated effectors Cas12 and Cas9, respectively. Recent studies have demonstrated that TnpB and IscB function as RNA-guided DNA endonucleases, but the broader biological role of this activity has remained enigmatic. Here we show that TnpB and IscB are essential to prevent permanent transposon loss as a consequence of the TnpA transposition mechanism. We selected a family of related insertion sequences from Geobacillus stearothermophilus that encode several TnpB and IscB orthologues, and showed that a single TnpA transposase was broadly active for transposon mobilization. The donor joints formed upon religation of transposon-flanking sequences were efficiently targeted for cleavage by RNA-guided TnpB and IscB nucleases, and co-expression of TnpB and TnpA led to substantially greater transposon retention relative to conditions in which TnpA was expressed alone. Notably, TnpA and TnpB also stimulated recombination frequencies, surpassing rates observed with TnpB alone. Collectively, this study reveals that RNA-guided DNA cleavage arose as a primal biochemical activity to bias the selfish inheritance and spread of transposable elements, which was later co-opted during the evolution of CRISPR-Cas adaptive immunity for antiviral defence.
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http://dx.doi.org/10.1038/s41586-023-06597-1 | DOI Listing |
J Glob Antimicrob Resist
December 2024
Antimicrobial Resistance Research Center, National Institute of Infectious Diseases, Tokyo, Japan.
Objectives: KLUC β-lactamase is a minor extended-spectrum β-lactamase (ESBL) derived from chromosome-encoded cefotaximase in Kluyvera cryocrescens. This study aimed to characterize the genetic context of KLUC-3-producing Escherichia coli and bla-harboring plasmids and assess nosocomial transmission.
Methods: In a national genomic surveillance conducted in 2019 and 2020, KLUC-3-producing E.
Vet J
December 2024
College of Animal Science and Technology, Northwest A&F University, Yangling 712100, China. Electronic address:
Immunoglobulins are important components of humoral immunity and play a crucial role in protecting the body from external antigens. The Arctic fox is an important member of furbearer farming, but due to the lack of research on the immune system of the Arctic fox, animal welfare regarding Arctic fox farming has still not received enough attention. In this study, we used the Arctic fox as a research subject, described the gene locus structure of the Arctic fox immunoglobulin germline by genome comparison, and analysed the mechanism of expression diversity of the antibody pool of the Arctic fox by rapid amplification of cDNA 5' ends and high-throughput sequencing.
View Article and Find Full Text PDFNat Commun
December 2024
Department of Microbiology and Molecular Genetics, University of Vermont, Burlington, VT, 05405, USA.
8-oxoguanine (8-oxoG) is a common oxidative DNA lesion that causes G > T substitutions. Determinants of local and regional differences in 8-oxoG-induced mutability across genomes are currently unknown. Here, we show DNA oxidation induces G > T substitutions and insertion/deletion (INDEL) mutations in human cells and cancers.
View Article and Find Full Text PDFmBio
December 2024
Division of Infectious Diseases, Brigham and Women's Hospital, Boston, Massachusetts, USA.
Unlabelled: Bacteria have evolved diverse strategies to ensure survival under nutrient-limited conditions, where rapid energy generation is not achievable. Here, we performed a transposon insertion site sequencing loss-of-function screen to identify genes that promote pathogen fitness in stationary phase. We discovered that the aintenance of ipid symmetry (Mla) pathway, which is crucial for transferring phospholipids from the outer to the inner membrane, is critical for stationary phase fitness.
View Article and Find Full Text PDFUnlabelled: strain E264 ( E264) and close relatives stochastically duplicate a 208.6 kb region of chromosome I via RecA-dependent recombination between two nearly identical insertion sequence elements. Because homologous recombination occurs at a constant, low level, populations of E264 are always heterogeneous, but cells containing two or more copies of the region (Dup+) have an advantage, and hence predominate, during biofilm growth, while those with a single copy (Dup-) are favored during planktonic growth.
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