Browse
Submit Data
Databases
API
Help

Dataset Information

0 Views

0 Connections

0 Citations

0 Reanalyses

0 Downloads

Omics score: 0

The comprehensive interactomes of human adenosine RNA methylases and demethylases revealed by proximity labelling-mass spectrometry.

ABSTRACT: N6-methyladenosine (m6A) and N6,2′-O‐dimethyladenosine (m6Am) are two abundant modifications found in mRNAs and ncRNAs that can regulate multiple aspects of RNA biology and function mainly through regulation of interaction with specific RNA-binding proteins. Both modifications are linked to development, disease and stress response. To date three methylases and two demethylases have been identified that modify adenosines in mammalian (m)RNAs. Here we provide a comprehensive study of the in vivo protein-protein interactome of the key methyltransferases and demethylases using the BioID proximity biotinylation approach, which allows capturing of stable and short-lived interactions. Our results show that METTL3, METTL16, CAPAM, FTO and ALKBH5 occupy specific non overlapping protein interacting territories. Moreover, the individual interactomes help to understand the biological functions of these enzymes

INSTRUMENT(S): Orbitrap Fusion Lumos, LTQ Orbitrap Elite

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Cell Culture

SUBMITTER: Ranjani Ganji

LAB HEAD: Zbynek Zdrahal

PROVIDER: PXD021566 | Pride | 2021-11-02

REPOSITORIES: Pride

ACCESS DATA

Json Xml

Dataset's files

Source:

			Action	DRS
	BAIT1_Control_BirA_01.raw	Raw
	BAIT1_Control_BirA_02.raw	Raw
	BAIT1_Control_BirA_03.raw	Raw
	BAIT1_Control_BirA_04.raw	Raw
	BAIT1_Control_BirA_05.raw	Raw

Items per page:

1 - 5 of 74

Similar Datasets

SLC15A4 functions as a metabolic guardian of immune cells by controlling mTORC1 and AMPK

Project description:Solute carrier family 15 member A4 (SLC15A4) is a lysosome-resident amino acid/oligopeptide transporter preferentially expressed in immune cells, and plays critical roles in the pathogenesis of lupus and colitis in murine model. SLC15A4 is required for TLR7/9- or NOD1-mediated inflammatory responses through mTORC1 regulation or transporting NOD1 ligand, TriDAP, respectively. To further understand the mechanisms of SLC15A4’s action in inflammatory responses, we applied proximity-dependent biotin identification experiment (BioID) to SLC15A4, and identified a panel of proteins including mTORC1 and its regulator that associate with and/or are proximate to SLC15A4 in vivo.

2021-08-11 | PXD020370 | Pride

Phosphoproteomics Identifies Dual-Site Phosphorylation in an Extended Basophilic Motif Regulating FILIP1-mediated Degradation of filamin-C FLNc d18-21 BioID experiment

Project description:FLNc, the muscle-specific isoform of the filamin family, is a multi-adaptor protein, comprising 1 amino-terminal actin-binding (ABD) domain followed by 24 immunoglobulin-like (Ig) domains. While FLNc can form homodimers via the last Ig-like domain and thus function as an actin-crosslinker like the other filamins, it features a unique insertion of 82 amino acids (aa) in domain 20. This insert was not only shown to mediate the interaction to several FLNc binding partners, but also to contain an Akt-mediated phosphorylation site at S2234 of mouse FLNc (mFLNc). To reveal novel proteins in the nano-environment of FLNc within myotubes under mild electrical pulse stimulation conditions, we applied a quantitative BioID appraoch.

2021-07-12 | PXD009159 | Pride

Fasnall induces atpically transient Stress Granules

Project description:Stress Granule formation has been linked to the resistance of some cancer cells to chemotherapeutic intervention. A number of studies have proposed that certain anti-tumor compounds promote cancer cell survival by inducing Stress Granules formation, leading to increased cellular fitness and apoptosis avoidance. Here we show that a potent fatty acid synthase inhibitor, fasnall, known for its anti-tumor capabilities, triggers the formation of non-canonical Stress Granules with a faster clearing and internal dynamics kinetics independently of fatty acid synthase inhibition. PABPC1 BioID interactomes in fasnall, and arsenite conditions were obtained.

2021-09-09 | PXD021229 | Pride

PABPC1 stress granule interactome identified by BioID

Project description:Identification of Stress Granule interacting proteins by BioID (Roux et al., 2012) using PABPC1-BirA(R118G)-GFP construct

2022-02-17 | PXD025334 | Pride

BioID screening experiment to map proteins linked to PHYHD1

Project description:PHYHD1 is a 2-oxoglutarate-dependent dioxygenase. To shed light on the cellular function of human PHYHD1, we performed a BioID screening experiment to map proteins linked to PHYHD1 functionally. For that, we used HEK293T cells transiently expressing either BirA* or BirA*-tagged PHYHD1 in the absence or presence of biotin.

2023-05-27 | PXD041792 | Pride

The Kdm5c histone demethylase controls enhancer and promoter function.

Project description:Here, we show that the Kdm5c/Smcx member of the Jarid1 family of H3K4 demethylases is recruited to both enhancer and core promoter elements in ES and neuronal progenitor cells (NPC). Knockdown of Kdm5c deregulates transcription via a local increase in H3K4me3. While at core promoters the function of Kdm5c is to restrict transcription, loss of Kdm5c impairs enhancer function. Remarkably, an impaired enhancer function activates promoter activity from Kdm5c-bound intergenic regions. Our results demonstrate that the Kdm5c demethylase plays a crucial role in the functional identity and discrimination of enhancers and core promoters. We speculate that this is related to recruitment of H3K4me3 binders like the TFIID and NURF complexes6-8. Providing functional identity to genomic regions through balancing enzymes that deposit and remove histone modifications may prove to be a general epigenetic mechanism for the functional indexing of eukaryotic genomes. Examination of the KDM5C binding sites in mouse embryonic stem cells and in neuronal progenitor cells. Effect of KDM5C knock down on H3K4me3 and H3K4me1 levels and gene expression.

2013-04-01 | E-GEOD-34975 | biostudies-arrayexpress

RNF43 truncations trap CK1 to drive niche-independent self-renewal in cancer

Project description:Wnt/b-catenin signaling is a primary pathway for stem cell maintenance during tissue renewal and a frequent target for mutations in cancer. Impaired Wnt receptor endocytosis due to loss of the ubiquitin ligase RNF43 gives rise to Wnt-hypersensitive tumors that are susceptible to anti-Wnt-based therapy. Contrary to this paradigm, we identify a class of RNF43 truncating cancer mutations that induce b-catenin-mediated transcription, despite exhibiting retained Wnt receptor downregulation. These mutations interfere with an ubiquitin-independent suppressor role of the RNF43 cytosolic tail that involves casein kinase-1 (CK1) binding and phosphorylation. Mechanistically, truncated RNF43 variants trap CK1 at the plasma membrane, thereby preventing b-catenin turnover and propelling ligandindependent target gene transcription. Gene editing of human colon stem cells shows that RNF43 truncations cooperate with p53 loss to drive a niche-independent program for self-renewal and proliferation. Moreover, these RNF43 variants confer decreased sensitivity to anti-Wnt-based therapy. Our data demonstrate the relevance of studying patient-derived mutations for understanding disease mechanisms and improved applications of precision medicine.

2020-08-14 | PXD020478 | Pride

Identification of Stress Granule interactor proteins in yeast S. cerevisiae

Project description:We investigated the sub-cellular architecture of Stress Granules in the context of glucose starvation. In order to uncover transient and peripheral Stress Granule interactors, we optimized the proximity-dependent biotinylation (BioID) approach for use in yeast. This approach allowed us to visualize Stress Granule formation while simultaneously inducing the biotinylation of SG components and nearby proteins in live cells. We used two independent Stress Granule components Pub1 and Pab1.

2020-03-23 | PXD016988 | Pride

Distinct interactomes of ADAR1 nuclear and cytoplasmic protein isoforms and their responses to Interferon induction

Project description:Adenosine deaminase acting on RNA 1 (ADAR1), is an enzyme that catalyzes the conversion of adenosine to inosine in double-stranded RNA, a process critical for regulation of innate immune response and distinguishing between ‘self and non-self RNA’. It is expressed as two isoforms: nucleolar p110 and cytoplasmic, interferon (IFN)-inducible, p150. The interactome of the p110 under steady-state conditions is well-studied; however, less is known about the interactions of the p150 isoform, particularly during IFN response. To elucidate ADAR1's protein interactions during IFN stimulation, alongside steady-state conditions, three distinct methods of enrichment were used followed by liquid chromatography-tandem mass spectrometry (LC-MS/MS). These included: immunoprecipitation (IP) of endogenous ADAR1, IP of Strep II-tagged ADAR1, and proximity labeling using BioID. Individual ADAR1 isoforms (p110 and p150) and their respective dsRNA binding-deficient mutants were created to discern isoform-specific and dsRNA-dependent interactions. Altogether, our results reveal a comprehensive ADAR1 interaction map, identifying both known and novel partners, and highlighting the isoform-specific and dsRNA-binding-dependent nature of ADAR1 interactions. Under IFN stimulation, ADAR1's interaction spectrum encompasses viral replication inhibitors and LINE-1 regulators. Mimicking viral infection with HMW poly(I:C) changed the proximal network of proteins for both isoforms. Our findings provide new insights into ADAR1's roles and its dynamic during IFN response.

2024-12-05 | PXD049681 | Pride

A proximity-labeling proteomic approach to investigate invadopodia molecular landscape in breast cancer cells

Project description:Invadopodia constitute a specialized machinery required for invasive tumor cells to travel through basement membranes and the interstitial matrix, and thus represent a key feature of metastatic cells. Therefore a better understanding of invadopodia biology and characteristics might permit the identification of markers of metastatic potential. Substantial efforts have been made to characterize invadopodia molecular composition, dynamics or triggering stimuli. However, the full molecular identity of these structures is still missing due to the fact that the isolation and purification of these structures has been challenging. To fill this gap, we developed a non-hypothesis driven proteomic approach based on the innovative BioID proximity biotinylation approach using the invadopodia-specific protein Tks5/FISH (its long form, Tks5) fused to the biotin ligase BirA as bait. As control, we also generated cells expressing the short form of Tks5 (Tks5) unable to localize to invadopodia. After validation of our cell models, Tks5 close neighbors were identified by label-free mass spectrometry after pulldown of biotinylated proteins. Known invadopodia components were identified revealing the pertinence of our strategy. Furthermore, this strategy allowed us to identify novel Tks5 in cellulo partners that appear as potential new invadopodia components/regulators and potential markers of metastasis.

2020-04-24 | PXD015847 | Pride

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