Browse
Submit Data
Databases
API
Help

Dataset Information

0 Views

0 Connections

0 Citations

0 Reanalyses

0 Downloads

Omics score: 0

Functional viromic screens uncover regulatory RNA elements

ABSTRACT: The number of sequenced viral genomes has surged recently, presenting an opportunity to understand viral diversity and uncover unknown regulatory mechanisms. Here we conducted a screening of 30,367 viral segments from 143 species representing 96 genera and 37 families. Using a library of viral segments in 3′ UTR, we identified hundreds of elements impacting RNA abundance, translation, and nucleocytoplasmic distribution. To illustrate the power of this approach, we investigated K5, an element conserved in kobuviruses, and found its potent ability to enhance mRNA stability and translation in various contexts, including adeno-associated viral vectors and synthetic mRNAs. Moreover, we identified a previously uncharacterized protein, ZCCHC2, as a critical host factor for K5. ZCCHC2 recruits the terminal nucleotidyl transferase TENT4 to elongate poly(A) tails with mixed sequences, delaying deadenylation. This study provides a unique resource for virus and RNA research and highlights the potential of the virosphere for biological discoveries.

INSTRUMENT(S): Orbitrap Eclipse

ORGANISM(S): Homo Sapiens (human)

SUBMITTER: Jenny Seo

LAB HEAD: V. Narry Kim

PROVIDER: PXD041296 | Pride | 2023-06-23

REPOSITORIES: Pride

ACCESS DATA

Json Xml

Similar Datasets

Discovery of Widespread Host Protein Interactions with the Pre-Replicated Genome of CHIKV using VIR-CLASP

Project description:The pioneer interactions between incoming viral RNA genomes and host proteins are crucial to infection and immune response. Until now, the ability to study these events was lacking. We developed VIR-CLASP (VIRal Cross-Linking And Solid-phase Purification) to characterize the earliest interactions between viral RNA and cellular proteins. We investigated the infection of human cells using Chikungunya virus (CHIKV) and Influenza A virus and identified hundreds of direct RNA-protein interactions. Here, we validate the biological impact of three protein classes that bind CHIKV RNA within minutes of infection. We find CHIKV RNA binds and hijacks the lipid-modifying enzyme FASN for pro-viral activity. We show that CHIKV genomes are N6-methyladenosine modified and that YTHDF1 binds and suppresses its replication. Finally, we find that the innate immune DNA sensor IFI16 associates with CHIKV RNA, reducing viral replication and maturation. Our findings have direct applicability to the investigation of potentially all RNA viruses.

2020-05-11 | PXD015863 | Pride

Leptobotia elongata

Project description:Leptobotia elongate Leptobotia elongate (Cypriniformes: Cobitidae) microsatellite sequence

| PRJNA205461 | ENA

Leptobotia elongata

Project description:Leptobotia elongate Leptobotia elongate (Cypriniformes: Cobitidae) microsatellite sequence

| PRJNA205477 | ENA

Influenza viral RNA pulldown MS in THP-1 cells

Project description:Due to the RNA nature of their genomes, influenza viruses have to utilize many RNA-binding proteins (RBPs) of both viral and host origin, for their replication. To uncover the comprehensive vRNA-host protein interactions, we performed affinity purification coupled with mass spectrometry (AP-MS) analysis of influenza vRNA complexes. The eight vRNA segments of H7N9 were transcribed and individually labeled with biotin in vitro and incubated with IAV virus-infected THP-1 cells, and vRNA complexes were enriched streptavidin magnetic beads and analyzed by mass spectrometry

2021-03-25 | PXD020723 | Pride

Comparative_Interaction_Proteomics_GFP-tagged_MAGEC2_HEK293T

Project description:293T (Human embryonic kidney cells-ATCC CRL-3216) cells were plated at 500,000 cells per 10 cm dish and grown for 48 hours. Cells were then transfected with GFP-MAGEC1 WT or GFP-MAGEC1 L318D in a 3:1 ratio of PEI (polyethylenimine) to DNA. Cells were harvested after 48 hours and lysed in 50 mM Tris pH 7.5, 150 mM NaCl, 1% Triton X-100, protease inhibitor (cOmpleteâ„¢, Mini, EDTA-free (Roche)). Cells were incubated for 25 minutes on ice and then sonicated for 5 minutes at 4Â°C. Cells were centrifuged at 14,000 rpm for 20 minutes at 4 Â°C. 500 ug lysate was then added to GFP-TrapÂ® Agarose resin (ChromoTek) and samples were incubated rotating at 4Â°C for 2 hours. Following washing of the beads with 50 mM Tris pH 7.5, 150 mM NaCl, proteins were eluted by the addition of 2x Laemmli buffer and boiled for 10 minutes at 95 Â°C. Samples were spun down and the supernatants were sent for mass spectrometry analysis as described below. Experiments were conducted in triplicate.

2024-02-20 | PXD047849 | Pride

Existence of human-virus chimeric proteins generated during infection

Project description:Influenza A virus (IAV) is a threat to mankind because it generates yearly epidemics and poorly predictable sporadic pandemics with catastrophic potential. Influenza has a small RNA genome (~14 Kb) composed of 8 mini-chromosomes (segments). Segments encode both structural proteins and proteins expressed only during infection. Segments are constituted by a 5’UTR followed by a coding region and a 3’UTR. Transcription of IAV RNA into mRNA depends on host RNA Polymerase II, as the viral polymerase cleaves 5’ capped cellular nascent transcripts to be used as primers to initiate mRNA synthesis. We hypothesized that host nascent transcripts bearing AUG could generate upstream ORFs in the viral segments, a phenomenon that would depend on the translatability of the viral 5’UTRs. Using orthogonal datasets we report the existence of this mechanism, which generate host-virus chimeric proteins. We show that most segments encode proteins in this manner, expanding the proteome diversity of IAV in infected cells. Host-virus chimeric proteins are conserved across IAV strains, pointing to an evolutionary conservation of function achieved by sampling of the evolutionary space before gene fixation. Thus, we discover a mechanism that generate human-virus chimeric proteins during infection.

2020-06-25 | GSE128519 | GEO

Reovirus low-density particles package viral and cellular RNA

Project description:Packaging of segmented, double-stranded RNA viral genomes requires coordination of multiple viral proteins and RNA segments. For mammalian orthoreovirus (reovirus), evidence suggests either all ten or zero viral RNA segments are simultaneously packaged in a highly coordinated process hypothesized to exclude host RNA. Accordingly, reovirus generates genome-containing virions and “genomeless” top component particles. However, despite ostensibly lacking the genome, top component particles maintain a low level of infectivity. Whether reovirus particles can package host RNA is unknown. To gain insight into reovirus packaging potential and mechanisms, we employed next-generation RNA-sequencing to define the viral and host RNA content of purified reovirus virions and top component particles. Reovirus top component particles contained double-stranded viral RNA segments in similar proportions but at reduced levels compared to virions. Top component particles also were enriched for numerous host RNAs, especially short, non-polyadenylated transcripts, that differed by reovirus strain, independent of the viral polymerase. In contrast, virions were enriched for very few host RNAs. Collectively, these findings indicate that genome packaging into reovirus virions is exquisitely selective, while incorporation of host RNAs into top component particles is more promiscuous or differentially selective and may contribute to or result from inefficient viral RNA packaging.

2021-06-08 | GSE164270 | GEO

Transcriptional analysis of apoptotic cerebellar granule neurons following rescue by Gip, Sp, Pacap and Igf1

Project description:Primary cultures of Cerebellar Granule Neurons (CGNs) have been extensively utilized to examine the signal transduction mechanisms underlying neuronal apoptosis. We conducted whole-genome expression profiling to decipher the transcriptional program controlling the apoptotic/survival switch in cerebellar granule neurons (CGNs) following the induction of apoptosis by serum and potassium deprivation and their rescue by gastric inhibitory polypeptide (Gip), substance p (Sp), insulin-like growth factor-1 (Igf1) or pituitary adenylyl cyclase-activating polypeptide (Pacap). Our results reveal the transcriptional changes intersecting neuronal apoptosis and survival and form the basis for further functional analyses and pharmacological exploitation to identify neuroprotective drugs. After six days âin vitroâ (DIV), extracellular KCl of CGNs was shifted from 25 to 5 mM for neuronal apoptotic death induction. After two washes with serum-free BME containing 5 mM KCl, neurons were incubated with the same medium for 6 h (K5), while control neurons were incubated with serum free medium supplemented with 25 mM KCl (K25). K5 neurons were also treated with a maximal effective dose of Gip, Sp, Igf1 and Pacap. Four biological replicates (derived from the same litter) for each of the experimental conditions (K25, K5, K5 + Gip; K25, K5, K5 + Sp; K25, K5, K5 + Igf1; K25, K5, K5 + Pacap) were analyzed.

2016-07-03 | E-GEOD-67788 | biostudies-arrayexpress

Analysis of mRNA expression differences in the skin of Casp-8F/–K5-Cre and Casp-8F/+K5-Cre mice

Project description:This SuperSeries is composed of the following subset Series: GSE17344: Analysis of spatial mRNA expression differences in the skin of Casp-8F/–K5-Cre or Casp-8F/+K5-Cre mice GSE17345: Analysis of temporal mRNA expression changes in cultured keratinocytes from Casp-8F/–K5-Cre and Casp-8F/+K5-Cre mice Refer to individual Series

2009-07-29 | E-GEOD-17365 | biostudies-arrayexpress

The balance of Influenza A Virus segments is achieved by virus-derived small non-coding RNAs

Project description:Encoding only ten major proteins, Influenza A virus (IAV) gains access to both the cell and nucleus, replicates its eight genomic segments, then enters the cytoplasm to assemble and egress. This is achieved through an expansive network of interactions with the host and a coordinated production of viral components. Here we show that upon infection, primary transcription of the virus results in the accumulation of the splice product, Nuclear Export Protein (NEP), which associates with the viral polymerase leading to the synthesis of segment-specific small viral RNAs (svRNAs). Here we demonstrate that svRNA biology is responsible for coordinating segment-specific amplification and is predominantly required for amplification of the three larger polymerase segments. Generation of svRNA ambiguity results in predictable changes in segment-specific vRNA levels and up to a five log attenuation. Together, these data elucidate the molecular function of svRNAs as essential mediators responsible for maintaining a stoichiometric balance between genomic segments and establish these small RNAs as functional components of the virus.

2017-06-14 | GSE83289 | GEO

OmicsDI is part of the ELIXIR infrastructure

OmicsDI is an Elixir interoperability service. Learn more ›

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