Browse
Submit Data
Databases
API
Help

Dataset Information

0 Views

0 Connections

0 Citations

0 Reanalyses

0 Downloads

Omics score: 0

INO80E_HIRIP3_RNA_duplex_proteins_RAP_MS

ABSTRACT: MS data from the RAP-MS experiment performed for identyfication of proteins that interact with HIRIP3 and INO80E RNA duplex.

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Embryonic Cell, Cell Culture, Kidney

SUBMITTER: Natalia Ryczek

LAB HEAD: Izabela Makałowska

PROVIDER: PXD060587 | Pride | 2025-02-21

REPOSITORIES: Pride

ACCESS DATA

Json Xml

Dataset's files

Source:

			Action	DRS
	209133611_1.raw	Raw
	209133611_1.xml	Xml
	209133612_2.raw	Raw
	209133612_2.xml	Xml

Items per page:

1 - 4 of 4

Similar Datasets

RNA-RNA interactions enable specific targeting of noncoding RNAs to nascent pre-mRNAs and chromatin sites

Project description:We developed a general method based on RNA Antisense Purification (RAP) to identify the intermolecular RNA-RNA interactions of a target RNA (RAP-RNA). RAP-RNA identifies endogenous RNA-RNA complexes through in vivo crosslinking, RNA capture with antisense oligonucleotides, and high-throughput RNA sequencing. This approach provides a systematic view of other RNAs that interact with a target RNA, and furthermore can distinguish between direct and indirect RNA-RNA interactions through the use of crosslinking reagents with different reactivities with proteins and nucleic acids. We applied this method to numerous small and large noncoding RNAs, including U1 snRNA, Malat1 lncRNA, Xist lncRNA, U3 snoRNA, U17/Snora73a snoRNA and U12 snRNA. We examined the RNA and chromatin interactions of ncRNAs in mouse embryonic stem cells. We developed and applied three related protocols: RAP-RNA[AMT], RAP-RNA[FA], and RAP-RNA[FA-DSG]. In the RAP-RNA[AMT] protocol, we fixed direct RNA-RNA hybrids in mouse embryonic stem (ES) cells with 4'-aminomethyltrioxalen (AMT), a psoralen-derivative crosslinker; AMT generates inter-strand crosslinks between uridine bases in RNA but does not react with proteins. In the RAP-RNA[FA] protocol, we used a different crosslinking strategy to capture both direct and indirect RNA-RNA interactions: we fixed ES cells using formaldehyde (FA), which crosslinks protein-RNA and protein-protein interactions and thus can capture both indirect interactions as well as direct interactions that are caged or flanked by proteins. In the RAP-RNA[FA-DSG] protocol, we fixed with both FA and disuccinimidyl glutarate (DSG), a strong protein-protein crosslinker, to more efficiently capture RNAs linked indirectly through multiple protein intermediates.

2014-09-01 | E-GEOD-55914 | biostudies-arrayexpress

ncRNA DUBR RAP-MS in HCT116 cells

Project description:Non-coding RNAs (ncRNAs) are finely tuned cellular regulators important for human cell growth and cancer progression. DUBR (also known as linc00883) is a nuclear ncRNA first discovered in mice for its role in regulating myoblast differentiation through interactions with chromatin and DNA methyltransferases. High expression levels of this ncRNA are predictive of poor patient outcome in colon adenocarcinoma, suggesting that DUBR may be involved in controlling cancer growth. To elucidate its function, we used RAP-MS and RNA immunoprecipitation techniques which revealed its interaction with epigenetic maintenance proteins in the human colon cancer cell line HCT116. Further, ATAC-seq and RNA-seq were used to address its function in regulating the epigenome and transcriptome of HCT116 cells. Here we report that DUBR is a regulator of human colon cancer cell line HCT116 survival.

2025-05-07 | PXD057811 | Pride

Integrator restraints paraspeckles assembly by promoting the 3’-end processing of the polyadenylated lncRNA NEAT1_1

Project description:Alternative 3’-end RNA processing provides an important source of transcriptome diversification, which impacts various biological processes and the etiology of diseases, including cancer. Here, we focused on a transcript isoform switch that leads to the readthrough expression of the long noncoding RNA NEAT1_2, at the expense of the shorter polyadenylated transcript NEAT1_1. Expression of NEAT1_2 is required for the formation of paraspeckles (PS), nuclear bodies that protect cancer cells from oncogene-induced replication stress and chemotherapy. Searching for proteins that interact with endogenous NEAT1 by RNA Affinity Purification coupled to mass spectrometry (RAP-MS), we identified factors involved in the 3’-end processing of polyadenylated (pA+) RNA as well as several components of the Integrator complex, known to process the 3’-ends of RNAs lacking polyadenylation sites. Unexpectedly, genetic perturbation experiments established that Integrator restraints NEAT1_2 expression, and thereby PS assembly, by promoting the 3’-end processing of pA+ NEAT1_1. Consistently, low expression levels of several Integrator subunits correlated with poorer prognosis of cancer patients exposed to chemotherapeutics. Our study identifies Integrator as a key regulator of PS biogenesis and highlights a previously unrecognized ability of the complex to process pA+ RNA. The data also establish a link between Integrator, cancer biology and chemosensitivity, which may be exploited therapeutically.

2020-10-14 | PXD015158 | Pride

Proteomic Analysis of RAP (RNA Antisense Purification) isolated RNPs (RiboNucleotide Particles) harboring the novel lncRNA mimi

Project description:In neurons, mRNAs and associated RNA-binding proteins assemble into ribonucleoprotein (RNP) granules essential to regulate mRNA trafficking, local translation, and turnover. Dysregulation of RNA-protein condensation can disturb synaptic plasticity. We report that the novel lncRNA mimi is a constitutive and essential component of large cytoplasmic condensates (RNP granules) in fly neurons. In order to identify direct mimi RNA binders we employ RAP assisted purification of mimi RNPs subsequent to UV-crosslinking of adult fly brain tissue (non UV irradiated samples serve as control). Applying relative Max Quant LFQ quantification (Max LFQ) we carry out a differential proteomic analysis of mimi RNP complexes in UV-irradiated versus non irradiated fly brains.

2022-10-12 | PXD034457 | Pride

Phosphoproteomic studies to identify targets and substrates of Protein kinase PfPK2 in human malaria parasite Plasmodium falciparum

Project description:The purpose of this project was to identify targets and substrates of Protein kinase PfPK2 in malaria parasite P. falciparum. These studies will help understand the mechanisms via which PfPK2, which is indispensable for the parasite survival, regulates parasite development. For this purpose, PfPK2-loxP parasite line was generated for conditional knock down of PfPK2 using rapamycin (RAP), which was used to induce the dimerization of active Cre-recombinase resulting in the depletion of PfPK2.

2023-10-18 | PXD042264 | Pride

Identification of proteins associated with the human mitochondrial degradosome

Project description:The human mitochondrial degradosome is a complex of the ribonuclease PNPase (Q8TCS8, encoded by PNPT1 gene) and RNA helicase SUV3 (Q8IYB8, encoded by SUPV3L1 gene). The aim of the project was to identify proteins which co-purify with both subunits of the degradosome. To this end we used human 293 cells stably expressing hSuv3 or PNPase with a C-terminal TAP tag or TAP tag fused to a mitochondria targeting sequence (control bait) (cell lines are described in details in (PMID: 19864255). Mitochondria were isolated from the cells, lysed and the protein extracts were subjected to affinity chromatography. Co-purified proteins were identified by mass spectrometry and their amounts were quantified by a label-free approach (MaxQuant). These experiments are part of the project “Dedicated surveillance mechanism controls G-quadruplex forming non-coding RNAs in human mitochondria”.

2018-07-06 | PXD009826 | Pride

Larch microsporocytes cytosplasmic fraction

Project description:Proteomic analysis of different stages of larch microsporocytes middle diplotene.

2022-06-09 | PXD032074 | Pride

The trypanosome Variant Surface Glycoprotein mRNA is stabilized by an essential unconventional RNA-binding protein

Project description:Salivarian trypanosomes cause human sleeping sickness and economically important livestock diseases, transmitted by Tsetse flies and replicating extracellularly throughout the life cycle. The surface of "bloodstream forms", which live within mammals, is coated by a monolayer of a Variant Surface Glycoprotein (VSG). Switching of the expressed VSG gene enables the parasites to evade adaptive immunity. Up until now, the mechanism by which VSG mRNA stability is maintained was unknown. To identify proteins specifically interacting with VSG mRNA (and alpha tubulin as control), we performed RNA-antisense purifications (RAP) following instructions published by McHugh et al. 2018 with some modifications.

2021-04-14 | PXD021772 | Pride

RNA antisnse puriification of Tau exon 10 pre-mRNA

Project description:In this study, we demonstrated the use of the RNA Antisense Purification by Mass Spectrometry (RAP-MS) method to identify proteins involved in specific alternative splicing events. Specifically, we applied the RAP-MS method to identify proteins that interact with the Tau pre-mRNA around its exon 10 region.

2022-02-16 | PXD023523 | Pride

The transcriptome of anthers of rice gamyb mutant expressing various GAMYB homologs was measured at the microspore stage.

Project description:To confirm the function of the GAMYB homologs for Selaginella moellendorffii and Physcomitrella patens in rice cells, we examined the expression profile in the anthers of transgenic plants, which OsGAMYB, SmGAMYB, or PpGAMYB2 is expressed under the control of the rice GAMYB promoter in rice gamy mutant. Expression profiling was conducted using rice 4x44K microarray RAP-DB (G2519F#15241). The microarray was used to profile gene expression in OsGAMYB:OsGAMYB, OsGAMYB:SmGAMYB, OsGAMYB:PpGAMYB2 and vector transgenic plants. To assess the reproducibility of the microarray analysis, each microarray experiment was performed twice using RNA samples that had been isolated independently (i.e., two biological replicates), and included a dye-swapping test as a technical replicate for each biological replicate.

2011-11-07 | E-GEOD-32652 | biostudies-arrayexpress

OmicsDI is part of the ELIXIR infrastructure

OmicsDI is an Elixir interoperability service. Learn more ›

Tweets

OmicsDI Databases

PRIDE
PeptideAtlas
MassIVE
JPOST Repository
Physiome Model Repository

EGA
EVA
ENA
LINCS
PAXDB
Cell Collective

MetaboLights
Metabolomics Workbench
MetabolomeExpress
GNPS
BioModels
FAIRDOMHub

ArrayExpress
dbGaP
ExpressionAtlas
GEO
NODE

Information

Databases
Help
API
Contact us
Code on GitHub
Terms of use
Submit Data