Mapping PT modifications in bacteria
Ontology highlight
ABSTRACT:
PROVIDER: PRJNA560558 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR9988660_1.fastq.gz | Fastqsanger.gz | |||
| SRR9988660_2.fastq.gz | Fastqsanger.gz | |||
| SRR9988661_1.fastq.gz | Fastqsanger.gz | |||
| SRR9988661_2.fastq.gz | Fastqsanger.gz | |||
| SRR9988662_1.fastq.gz | Fastqsanger.gz |
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Project description:One microbial organism always contains multiple DNA modification systems as defense strategies or epigenetic controls. Here, we describe crosstalk between two distinct DNA modifications with different target sequence motif: DNA phosphorothioation (PT, GPSAAC/GPSTTC by DndACDE proteins) and N6-methyl-adenine methylation (m6A, G6mATC by Dam protein). By using a newly developed DNA modification sequencing method PT nick-seq, we found that half of PT modifications change from GAAC/GTTC to GATC sites after bacteria have lost Dam.
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Project description:Endogenous and exogenous chemical modifications in DNA have profound influences on genome function. We have developed a novel technology, Nick-seq, for mapping variety of DNA chemical modifications across the genomes at single-nucleotide resolution, by which we achieved quantitative profiling of single-strand breaks, phosphorothioate modifications, and oxidative DNA damage sites. This method is applicable for mapping of any DNA metabolism events that are involved in or can be converted, enzymatically or chemically, to DNA strand breaks such as DNA modifications, damages, genome replication, and chromatin structures.
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Project description:Nick-seq for singlenucleotide resolution genomic maps of DNA modifications and damage:DNA PT modification mapping
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Project description:Endogenous and exogenous chemical modifications in DNA have profound influences on genome function. We have developed a novel technology, Nick-seq, for mapping variety of DNA chemical modifications across the genomes at single-nucleotide resolution, by which we achieved quantitative profiling of single-strand breaks, phosphorothioate modifications, and oxidative DNA damage sites. This method is applicable for mapping of any DNA metabolism events that are involved in or can be converted, enzymatically or chemically, to DNA strand breaks such as DNA modifications, damages, genome replication, and chromatin structures.
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Project description:Pt-ttpy (tolyl terpyridin-Pt complex) covalently binds to G-quadruplex (G4) structures in vitro and to telomeres in cellulo via its Pt moiety. Here, we identified its targets in the human genome, in comparison to Pt-tpy, its derivative without G4 affinity, and cisplatin. Pt-ttpy, but not Pt-tpy, induces the release of the shelterin protein TRF2 from telomeres concomitantly to the formation of DNA damage foci at telomeres but also at other chromosomal locations. -H2AX chromatin immunoprecipitation (ChIP-seq) after treatment with Pt-ttpy or cisplatin revealed accumulation in G- and A-rich tandemly repeated sequences, but not particularly in potential G4 forming sequences. Collectively, Pt-ttpy presents dual targeting efficiency on DNA, by inducing telomere dysfunction and genomic DNA damage at specific loci.
2021-05-04 | GSE171450 | GEO
Project description:RNA is subject to a multitude of different chemical modifications that collectively represent the epitranscriptome. Individual RNA modifications including N6-methyladenosine (m6A) on mRNA play essential roles in the posttranscriptional control of gene expression. Recent technological advances have enabled the transcriptome-wide mapping of certain RNA modifications, to reveal their broad relevance and characteristic distribution patterns. However, convenient methods that enable the simultaneous mapping of multiple different RNA marks within the same sample are generally lacking. Here we present EpiPlex RNA modification profiling, a bead-based proximity barcoding assay with sequencing readout that expands the scope of molecular recognition-based RNA modification detection to multiple targets, while providing relative quantification and enabling low RNA input. Measuring signal intensity against spike-in controls provides relative quantification, indicative of the RNA mod abundance at each locus. We report on changes in the modification status of HEK293T cells upon treatment with pharmacological inhibitors separately targeting METTL3, the dominant m6A writer enzyme, and the EIF4A3 component of the exon junction complex (EJC). The treatments resulted in decreased or increased m6A levels, respectively, without effect on inosine levels. Inhibiting the helicase activity of EIF4A3 and EIF4A3 knockdown both cause a significant increase of m6A sites near exon junctions, consistent with the previously reported role of EIF4A3 in shaping the m6A landscape. Thus, EpiPlex offers a reliable and convenient method for simultaneous mapping of multiple RNA modifications to facilitate epitranscriptome studies .
2024-10-17 | GSE278242 | GEO
Project description:Mapping diverse RNA modifications simulataneously by proximity barcode sequencing
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Project description:Endogenous and exogenous chemical modifications in DNA have profound influences on genome function. We have developed a novel technology, Nick-seq, for mapping variety of DNA chemical modifications across the genomes at single-nucleotide resolution, by which we achieved quantitative profiling of single-strand breaks, phosphorothioate modifications, and oxidative DNA damage sites. This method is applicable for mapping of any DNA metabolism events that are involved in or can be converted, enzymatically or chemically, to DNA strand breaks such as DNA modifications, damages, genome replication, and chromatin structures.
2020-05-14 | GSE138070 | GEO
Project description:Purpose: We investigate the evolutionary footprints of a bacteria-plasmid association consisting of Escherichia coli K-12 MG1655 and plasmid RP4 undergoing a long-term sub-MIC antibiotic stress. Methods: Bacterial mRNA profiles of evolved RP4-carrying strains (E:H:p) and ancestral RP4-carrying strains (A:H:p) were generated by deep sequencing on an Illumina Hiseq platform. The sequence reads that passed quality filters were analyzed by Burrows–Wheeler Aligner (BWA), followed by ANOVA (ANOVA) and TopHat followed by Cufflinks. qRT–PCR validation was performed using TaqMan and SYBR Green assays Results: Using an optimized data analysis workflow, we mapped about 15 million sequence reads of E:H:p and 12 million sequence reads of A:H:p to the E. coli MG1655 genome (GCF_000801205.1) and differential expressed genes were identified with TopHat workflow. RNA-seq data showed that approximately 15% of the transcripts showed differential expression between the E:H:p and A:H:p strains, with a fold change ≥1 and p value <0.005. Altered expression of 26 genes was confirmed with qRT–PCR, demonstrating the high degree of sensitivity of the RNA-seq method. Data analysis with bowtie and TopHat workflows provided complementary insights in transcriptome profiling. Conclusions: Our study showed the coevolved bacteria-plasmid pairs has colonization traits superior to the wild-type parent strain. Antibiotic stress was necessary for bacterial evolution and evolved strains mostly employed transcriptional modifications to reduce plasmid-related cost in evolutionary adaptations. Several genes related to chromosome-encoded efflux pumps were transcriptionally upregulated, while most plasmid-harboring genes were downregulated based on RNA gene sequencing. These transcriptional modifications endowed evolved strains with resistant phenotype modifications, including the enhanced bacterial growth and biofilm formation.
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