Browse
Submit Data
Databases
API
Help

Dataset Information

0 Views

0 Connections

0 Citations

0 Reanalyses

0 Downloads

Omics score: 0

The nuclear cap-binding complex regulates subcellular RNA processing and surveillance of coding and noncoding RNAs

ABSTRACT: RNA cleavage is essential for processing and regulating all classes of RNA. Current methods profiling genome-wide RNA cleavage are biased towards cytoplasmic events and ignore compartmentalized differences. Here, we couple subcellular RNA fractionation with degradome profiling to detect genome-wide nucleoplasm- and cytoplasm-enriched RNA cleavage in Arabidopsis thaliana. While messenger RNA (mRNA) cleavage dominated cytoplasmic fractions, we captured a diverse array of nucleoplasm-enriched RNA cleavage events. These included pre-mRNA cleavage and noncoding RNA processing, including for microRNAs, ribosomal RNAs, small nucleolar RNAs, enhancer-associated RNAs, and retrotransposon-derived RNA. Furthermore, our findings suggest that CAP-BINDING PROTEIN80/ABA HYPERSENSITIVE1 regulates mRNA surveillance within the perinuclear cytoplasm during the pioneer round of translation. Our data also emphasized its role in stabilizing nucleoplasmic RNAs (e.g. mRNA-associated antisense RNAs) and affecting cytoplasmic mRNA cleavage. Overall, our results highlight the diversity of compartmentalized RNA cleavage and reveal that the nuclear cap-binding complex has numerous functions in subcellular RNA processing and surveillance.

ORGANISM(S): Arabidopsis thaliana

PROVIDER: GSE291993 | GEO | 2025/05/09

REPOSITORIES: GEO

ACCESS DATA

Json Xml

Dataset's files

Source:

			Action	DRS
		Other

Items per page:

1 - 1 of 1

Similar Datasets

DEAD-box ATPase Dbp2 mediates mRNA release after 3’-end formation

Project description:mRNA biogenesis in the eukaryotic nucleus is a highly complex process. The numerous RNA processing steps are spatially and temporally coordinated to ensure that only fully processed transcripts are released into the nucleoplasm for export from the nucleus. Here, we explore the hypothesis that fission yeast Dbp2, a ribonucleoprotein complex (RNP) remodelling ATPase of the DEAD-box family, is involved in a RNP assembly checkpoint at the 3’-end of genes that is coupled to the release of the 3’-end processing complex after polyadenylation. We show that Dbp2 interacts with the cleavage and polyadenylation complex (CPAC) and localizes to cleavage bodies, which are enriched for 3’-end processing factors and proteins involved in nuclear RNA surveillance. Upon loss of Dbp2, 3’-processed, polyadenylated RNAs accumulate on chromatin and in cleavage bodies, which is accompanied by a depletion of CPAC components from the soluble pool. Under these conditions, cells display an increased likelihood to skip polyadenylation sites as well as delayed transcription termination, suggesting that the availability of CPAC components is insufficient to maintain normal levels of 3’-end processing. Our data is consistent with a model in which Dbp2 is involved in an mRNP remodelling checkpoint that licenses RNA export and is coupled to CPAC release.

2024-07-04 | PXD048560 | Pride

CBC-ARS2 stimulates 3'-end maturation of multiple RNA families and favors cap-proximal processing

Project description:The nuclear cap-binding complex (CBC) stimulates multiple steps in several RNA maturation pathways, but how it functions in humans is incompletely understood. For small capped RNAs like pre-snRNAs, the CBC recruits PHAX. Here, we identify the CBCAP complex, composed of CBC, Ars2 and PHAX, and show that both CBCAP and CBC-Ars2 complexes can be reconstituted from recombinant proteins. Ars2 stimulates PHAX binding to the CBC as well as snRNAs 3'-end processing, hereby coupling maturation with export. In vivo, CBC and Ars2 bind similar capped non-coding and coding RNAs, and stimulate their 3’-end processing. Strongest effects are displayed for cap-proximal polyadenylation sites, thereby favoring premature transcription termination. Ars2 functions in part through the mRNA 3’-end cleavage factor CLP1, which binds RNA Polymerase II through PCF11. Ars2 is thus a major CBC effector that stimulates functional and cryptic 3'-end processing sites.

2013-11-08 | GSE52131 | GEO

CBC-ARS2 stimulates 3'-end maturation of multiple RNA families and favors cap-proximal processing

Project description:The nuclear cap-binding complex (CBC) stimulates multiple steps in several RNA maturation pathways, but how it functions in humans is incompletely understood. For small capped RNAs like pre-snRNAs, the CBC recruits PHAX. Here, we identify the CBCAP complex, composed of CBC, Ars2 and PHAX, and show that both CBCAP and CBC-Ars2 complexes can be reconstituted from recombinant proteins. Ars2 stimulates PHAX binding to the CBC as well as snRNAs 3'-end processing, hereby coupling maturation with export. In vivo, CBC and Ars2 bind similar capped non-coding and coding RNAs, and stimulate their 3M-bM-^@M-^Y-end processing. Strongest effects are displayed for cap-proximal polyadenylation sites, thereby favoring premature transcription termination. Ars2 functions in part through the mRNA 3M-bM-^@M-^Y-end cleavage factor CLP1, which binds RNA Polymerase II through PCF11. Ars2 is thus a major CBC effector that stimulates functional and cryptic 3'-end processing sites. In total 12 samples; 4 control IP, 2 Flag- Ars2 IP and 2 Flag-CBP20 IP; 2 control FFL siRNA and 2 siRNA Ars2

2013-11-08 | E-GEOD-52131 | biostudies-arrayexpress

hCLE/RTRAF-HSPC117-DDX1-FAM98B: A new cap-binding complex that activates mRNA translation

Project description:hCLE/C14orf166/RTRAF, DDX1 and HSPC117 are components of cytoplasmic mRNA-transporting granules kinesin-associated in dendrites. They have also been found in cytoplasmic ribosome-containing RNA granules that transport specific mRNAs halted for translation until specific neuronal signals renders them accessible to the translation machinery. hCLE associates to DDX1, HSPC117 and FAM98B in HEK293T cells and all four proteins bind to cap analog-containing resins. Competition and elution experiments indicate that binding of hCLE complex to cap resins is independent of eIF4E; the cap-binding factor needed for translation. Purified hCLE free of its associated proteins binds cap with low affinity suggesting that its interacting proteins modulate its cap association. hCLE silencing reduces hCLE accumulation and that of its interacting proteins and decreases mRNA translation. hCLE-associated RNAs have been isolated and sequenced; RNAs involved in mRNA translation are specifically associated. The data suggest a positive role of hCLE complex modulating mRNA translation.

2019-02-18 | GSE84824 | GEO

The nuclear cap-binding complex regulates subcellular RNA processing and surveillance of coding and noncoding RNAs

Project description:The nuclear cap-binding complex regulates subcellular RNA processing and surveillance of coding and noncoding RNAs

| PRJNA1236297 | ENA

Transcriptome-Wide Analysis of the 5' Cap Status of RNA Using 5' Monophosphate-Dependent Exonuclease Digestion and RNA Sequencing

Project description:Eukaryotic mRNAs carry an N7-methylguanosine (m7G) cap structure at their 5' extremity, which protects them from the degradation by 5'-3' exoribonucleases and plays a pivotal role in mRNA metabolism, promoting splicing, nuclear export, and translation. Decapping, the enzymatic process that removes this structure, is a key event during cytoplasmic mRNA 5'-3' decay, leading to the degradation of the transcript body by Xrn1. In this chapter, we describe a procedure to assess the cap status of RNA at the transcriptome level. It is based on a treatment of total RNA extracts with a 5' monophosphate-dependent exonuclease, which like Xrn1 specifically degrades decapped RNAs harboring 5' monophosphate extremities, but not RNAs with intact m7G cap. The digested RNAs are then analyzed by RNA sequencing.

2024-11-14 | GSE203280 | GEO

Proximity-CLIP provides a snapshot of occupied cis-acting elements on RNA in different subcellular compartments on a transcriptome-wide scale

Project description:Many cellular RNAs localize to specific subcellular compartments. Currently, methods to systematically study subcellular RNA localization are limited and lagging behind proteomic approaches. Here, we combined APEX2-mediated proximity biotinylation of proteins with PAR-CLIP to simultaneously profile the proteome and the transcriptome bound by RNA binding proteins in any given subcellular compartment. Our approach is fractionation-independent and does not rely on additional RNA manipulation and labeling steps, thus making it easy to apply. Furthermore, it enables to study the locali-zation of RNA processing intermediates, as well as the identification of regulatory RNA cis-acting elements occupied in different cellular compartments. In a proof-of-concept study we studied RNA and protein localization in the nucleus, cytoplasm and at cell-cell interfaces using Proximity-CLIP. These experiments revealed among other in-sights frequent transcriptional readthrough continuing for several kilobases down-stream of the canonical cleavage and polyadenylation site, a differential binding pat-tern of nuclear and cytoplasmic mRNAs, as well as the localization of mRNAs contain-ing 3’UTR CUG sequence elements at cell-cell interfaces, of which many encode regulatory proteins.

2018-11-26 | GSE110380 | GEO

Multi-condition affinity capture/MS analysis of LAP-tagged human NCBP1, NCBP2, and NCBP3

Project description:A 5′, 7-methylguanosine cap is a quintessential feature of RNA polymerase II-transcribed RNAs, and thus a textbook aspect of co-transcriptional pre-mRNA processing. The cap is bound by the cap-binding complex (CBC), canonically consisting of nuclear cap-binding proteins 1 and 2 (NCBP1/2). Recently, NCBP3 has been proposed to form an alternative, non-canonical CBC, together with NCBP1. NCBP3 has also been shown to interact with the canonical CBC along with the protein SRRT (aka ARS2), in a manner that is mutually exclusive with the RNA export factor, PHAX. Taken together, ambiguities and missing information in the bona fide physiological protein-protein associations of NCBP3 persist. In an effort to clarify the compositions of NCBP1-, 2-, and 3-related macromolecular assemblies, including their intersections and differences, we have applied our recently developed interactome screening platform (PMID: 25938370). Here the experimental design and data processing have been modified and updated to identify interactome differences between targets of affinity capture under a wide range of experimental conditions, followed by label-free quantitative mass spectrometry.

2020-11-09 | PXD016038 | Pride

RNA-dependent RNA polymerase sequence specificities of capsnatching viruses are tailored to aid viral replication

Project description:Influenza A virus (IAV) is a major respiratory pathogen that causes severe illness and mortality worldwide. IAV critically depends on host factors for multiple functions that are not encoded by the viral genome. Notably, the IAV RNA-dependent RNA polymerase (RdRP) complex lacks intrinsic 7-methylguanosine (m7G) capping activity and instead “snatches” the 5’m7G caps and the adjacent 10-15nts of host transcripts to prime viral mRNA production, allowing viral transcripts to be recognized by the host translation machinery. We developed a new approach to simultaneously assay the 5’ ends of host and viral transcripts in infected cells and identify the host capsnatch origins for viral mRNAs. We used this technique to study the progression of IAV infection in nuclear and cytoplasmic compartiments and determined that the RdRP complex has specific sequence preferences that results in preferential capsnatching and avoidance of sub-populations of host RNAs, which can be reverted by the addition of a cap-snatch inhibitor. Notably, genes avoided by CapSnatch Seq have biological functions enriched for mRNA processing, translation, and viral replication. This selectivity and functional enrichment of avoided genes is conserved among capsnatching viruses including Influenza B Virus and Lymphocytic Choriomeningitis Virus, despite differences in RdRP domains and subcellular locations of RNA substrates. This suggests that this preference convergently arose as a viral strategy to allow cap-snatching viruses to avoid targeting genes necessary for its own replication.

2017-08-09 | GSE85474 | GEO

Analysis of Pro-Inflammatory Gene Activation and RNA Processing by RNAseq of Nascent Transcripts

Project description:We examined pro-inflammatory gene activation in activated murine macrophages by performing RNA-Seq with fractionated chromatin-associated, nucleoplasmic, and cytoplasmic transcripts. This experimental strategy allowed a global, high-resolution analysis of the transcriptional regulation of diverse classes of co-expressed genes, with a direct comparison to transcript processing and the transit of RNA from the chromatin to the nucleoplasm and the cytoplasm. The results provide quantitative insights into the relationship between transcription and distinct promoter and chromatin properties, as well as genome-scale insights into RNA processing and dynamics. RNA-seq of subcelluar fractions from five stimulation time-points

2012-05-02 | E-GEOD-32916 | 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