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Human proteins bound to RNA/DNA hybrid

ABSTRACT: Human proteins bound to RNA/DNA hybrid

PROVIDER: PRJNA482858 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR10143746.fastq.gz	Fastqsanger.gz
	SRR10143747_1.fastq.gz	Fastqsanger.gz
	SRR10143747_2.fastq.gz	Fastqsanger.gz
	SRR7592276.fastq.gz	Fastqsanger.gz
	SRR7592278.fastq.gz	Fastqsanger.gz

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Human proteins bound to RNA/DNA hybrid

Project description:RNA/DNA hybrids form when RNA hybridizes with its template DNA generating a three-stranded structure known as the R-loop. Knowledge of how they form and resolve, and their functional roles are limited. Here, we identified proteins that bind to RNA/DNA hybrids.

2018-08-03 | GSE117671 | GEO

Massive sequencing of hybrid DNA obtained by DNA:RNA immunoprecipitation (DRIP) in extracellular microvesicles (MV) produced by HeLa cells (using S9.6 antibody)

Project description:Hybrid (Bound) and non-hybrid (Unbound) DNA from Hela EV was isolated using DRIP procedure as follows. Between 1-5μg of nucleic acid DNA from EVs (DNase0 treated) was incubated overnight with 1μg of anti-DNA-RNA Hybrid (S9.6) antibody (KeraFast) under rotation at 4°C in 500μl of binding buffer (10mM NaPO4 pH 7.0, 140 mM NaCl, 0.05% Triton X-100). The following day, 25μl of A/G magnetic beads (Pierce™) were washed twice in 500μl of binding buffer (30' min each wash) and added for 2h at room temperature under rotation to the S9.6+nucleic acid complex. The nucleic acids bound to the S9.6 antibody (Hybrid fraction or Bound) were separated from those unbound (Non-Hybrid or Unbound) by using a magnetic rack (which captured the Bead+Hybrid fraction). 500μl of the unbound fraction (bead-free) were transferred into a new 1.5ml tube and collected for further analysis. The Bead+Hybrid fraction was washed twice in 250μl binding buffer for 15' min at room temperature under rotation. The hybrids were then eluted with 250μl of elution buffer (50mM TRIS pH 8.0, 10mM EDTA, 0.5% SDS) for 15' min at room temperature under rotation. The elution step was repeated twice to make sure that all the hybrid molecules were collected. As a control of all there experiments to make sure, we were isolating and precipitating Hybrid DNA:RNA molecules, samples were also pre-treated with RNaseH1 (RNaseH), which degrades the RNA filament only when present in the hybrid structure.

2023-03-30 | E-MTAB-9634 | biostudies-arrayexpress

bound proteins of DNA nanomaterials exposed to human serum

Project description:DNA aptamers and framework DNA nanostructures are emerging DNA materials with many appealing biological applications including biosensing, bioimaging, drug delivery etc. When placed in physiological fluids, they inevitably encounter biomolecules (majorly proteins) and form complexes that largely affects their biological fate. Nevertheless, little is known regarding the quantitative profile of proteins that adsorb to DNA aptamers and DNA nanostructures in biological environments, and there are no potent strategies to regulate protein profiles. Herein, we performed a proteomic analysis to profile proteins that bind to DNA aptamers (Sgc8c and SYLC3) and nanostructures (a tetrahedral DNA nanostructure and a DNA origami rod) in human serum by using liquid chromatography mass spectrometry (LC-MS).

2021-05-15 | MSV000087433 | MassIVE

Investigation of RNA editing sites within bound regions of RNA-binding proteins

Project description:Investigation of RNA editing sites within bound regions of RNA-binding proteins

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Immunoprecipitation of RNA-DNA hybrid interacting proteins in Trypanosoma brucei reveals conserved and novel activities, including in host immune evasion by antigenic variation

Project description:RNA-DNA hybrids are widespread epigenetic features of genomes that provide a growing range of activities in transcription, chromatin and DNA replication and repair. Understanding of these diverse functions has been advanced by characterising the proteins that interact with the hybrids, with all such studies revealing hundreds of potential interactors. However, all interaction analyses to date have focused on mammalian cells, and so it is unclear if a similar spectrum of RNA-DNA hybrid interactors found in other eukaryotes, thus limiting our understanding of the conserved and lineage-specific activities linked to these genetic structures. The African trypanosome is a compelling organism in which to address these questions. As a divergent single-cell eukaryotic parasite of the Discoba grouping, Trypanosoma brucei displays substantial divergence in several aspects of core biology from its mammalian host and, unusually for a protist, has well-developed tools for molecular genetic analysis. For these reasons, we used DNA-RNA hybrid immunoprecipitation coupled with mass spectrometry to reveal 602 putative interactors in T. brucei mammal- or insect vector-infective stage cells. We show that the approach selects for a subset of the parasite proteome and reveals a range of predicted RNA-DNA hybrid associated activities, some overlapping with similar studies in mammals. We demonstrate that loss of three factors, two putative helicases and a RAD51 paralogue, impact on T. brucei nuclear RNA-DNA hybrid and DNA damage levels. Moreover, loss of each affects the operation of the crucial parasite immune survival mechanism of antigenic variation. Thus, our work reveals the broad range of activities contributed by RNA-DNA hybrids to T. brucei biology, including new functions in host immune evasion as well as many conserved with mammals, and so likely fundamental to eukaryotic genome function.

2024-01-26 | PXD042146 | Pride

DNA lesions can frequently precede DNA:RNA hybrid accumulation [RNA-seq]

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Intergenerational genomic DNA methylation patterns in mouse hybrid strains

Project description:Intergenerational genomic DNA methylation patterns in mouse hybrid strains

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DNA lesions can frequently precede DNA:RNA hybrid accumulation [RNA-seq]

Project description:While DNA:RNA hybrids contribute to multiple genomic transactions, their unscheduled formation is a recognized source of DNA lesions. Here, through a suite of systematic screens, we rather observed that a wide range of yeast mutant situations primarily triggering DNA damage actually leads to hybrid accumulation. Focusing on Okazaki fragment processing, we established that genic hybrids can actually form as a consequence of replication-born discontinuities such as unprocessed flaps or unligated Okazaki fragments. Strikingly, such “post-lesion” DNA:RNA hybrids neither detectably contribute to genetic instability, nor disturb gene expression, as opposed to “pre-lesion” hybrids formed upon defective mRNA biogenesis, e.g., THO complex mutants. Post-lesion hybrids similarly arise in distinct genomic instability situations, triggered by pharmacological or genetic manipulation of DNA-dependent processes, both in yeast and human cells. Altogether, our data establish that the accumulation of transcription-born DNA:RNA hybrids can occur as a consequence of various types of natural or pathological DNA lesions, yet do not necessarily aggravate their genotoxicity.

2025-02-06 | GSE265892 | GEO

Transposase assisted tagmentation of RNA/DNA hybrid duplexes

Project description:Transposase assisted tagmentation of RNA/DNA hybrid duplexes

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DNA lesions can frequently precede DNA:RNA hybrid accumulation [DRIP-seq]

Project description:DNA lesions can frequently precede DNA:RNA hybrid accumulation [DRIP-seq]

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