MS for RNA binding preteins in tapasin 3'UTR
Ontology highlight
ABSTRACT:
INSTRUMENT(S):
ORGANISM(S): Homo Sapiens (human)
TISSUE(S): Melanocyte, Skin
DISEASE(S): Melanoma
SUBMITTER: 
Yuan Wang
LAB HEAD: Yuan Wang
PROVIDER: PXD045056 | Pride | 2025-05-06
REPOSITORIES: Pride
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| 1_CHRO001.DAT | Other | |||
| 1_CHRO002.DAT | Other | |||
| 1_CHRO003.DAT | Other | |||
| 1_CHRO004.DAT | Other | |||
| 1_CHRO005.DAT | Other |
Items per page: 5 1 - 5 of 43 |
Publications
Unconventional microRNA role: Enhancing the human leukocyte antigen class I antigen processing pathway via interacting with a silencer.
Wang Yuan Y Lazaridou Maria-Filothei MF Kordaß Theresa T Massa Chiara C Vaxevanis Christoforos K CK Eichmüller Stefan S Seliger Barbara B
Clinical and translational medicine 20241001 10
Similar Datasets
Project description:microRNAs binding with 3'UTR of tapasin
| PRJNA1004588 | ENA
Project description:Background Dysregulation of major histocompatibility complex (MHC) class I antigen processing and presentation machinery (APM) components in the tumor as one main molecular mechanism of immune escape leading to deactivation of T cell immune surveillance could be due to post-transcriptional regulation via immune-modulatory microRNAs (miRNA). It is now well established from a variety of studies that several miRNAs could effectively modulate the expression of some MHC class I APM components in tumors. Tapasin is an important APM molecule involved in the association of MHC class I with transporter associated with antigen processing (TAP) and peptide loading. Since so far no detailed investigation of the posttranscriptional regulation of tapasin exists, the aim of this study is to identify and functionally characterize miRNAs targeting tapasin in melanoma. Methods Using miRNA trapping by RNA in vitro affinity purification (miTRAP) and in silico as well as small RNA sequencing, miRNAs will be identified, which bind to the 3'untranslated region (3' UTR) of tapasin. Dual luciferase assays will be performed to determine binding of the miRNA. In silico analysis was performed to predict the effect of miRNAs on the survival of melanoma patients in correlation to tapasin. RT-qPCR, Western blot, flow cytometry and other functional assays were performed after transfecting miRNA mimics in three melanoma cell lines. Results Using the combination strategy of miTRAP and RNA seq we identified miR-155-5p to bind to the 3’UTR of tapasin, which was further confirmed by in silico analysis and dual luciferase reporter assay. Transfection of miR-155-5p mimics demonstrated that miR-155-5p upregulate tapasin protein level, which was accompanied by an upregulation of the MHC class I (HLA-ABC) surface expression. Simultaneously, in several different types of cancer, including melanoma, the expression of miR-155-5p is significantly positively correlated with the patient's survival and HLA-A protein. Conclusion Our data revealed for the first time a positive role of miR-155-5p in the posttranscriptional regulation of tapasin in melanoma and provide further insights into the miR-155-5p-mediated induction of HLA-ABC surface expression. This might lead to a better T cell response, avoidance tumor cell escape, improvement of patients' survival and thus might be a potential therapeutic target.
2024-09-15 | GSE240665 | GEO
Project description:Nrf2 transcript factor plays an important role in cellular defense against oxidative stress due to its control for the expression of antioxidant and detoxification genes. We have found that Nrf2 undergoes de novo protein translation when mammalian cells encounter oxidative stress. Here, we report the discovery of DHX9 as a binding protein for Nrf2 5’UTR. H2O2 treatment causes dose- or time- dependent increases of DHX9 binding to Nrf2 5’UTR, in parallel with elevation of Nrf2 protein. Inhibiting DHX9 expression with siRNA or its activity with YK-4-279 blocked H2O2 from inducing Nrf2 mRNA recruitment to the ribosomes or Nrf2 protein elevation. As a nuclear protein, DHX9 was found to increase its abundance in the cytosol with oxidative stress. An increase of DHX9 was detected in total ribosomal collections in cells treated with H2O2, most significantly with 100 uM H2O2, and at 60 mins. Ribosomal fractionation revealed an increase of DHX9 protein at 43/48S and 80S fractions in H2O2 treated cells. H2O2 treatment caused an RNA dependent increase of DHX9 interaction with eIF3 The binding of DHX9 to Nrf2 5’UTR probe was enhanced by H2O2 treated cells or by deducting the length of Nrf2 5’UTR, suggesting a negative element normally prohibiting DHX9 binding to Nrf2 5’UTR. RNase digestion enhanced DHX9 association with the ribosomes. Our data have revealed a novel mechanism of de novo Nrf2 protein translation under oxidative stress involving DHX9 binding to Nrf2 5’UTR, perhaps via removal of a negative RNA element, to recruit 43S pre-initiation complex for translation initiation.
2025-06-23 | PXD055838 | Pride
Project description:mTOR regulates mRNA translation. Whereas ribosome-profiling suggested that mTOR exclusively stimulates translation of TOP (containing a 5â-terminal oligopyrimidine [5âTOP] motif) and TOP-like mRNAs, polysome-profiling implied that mTOR also modulates translation of non-TOP mRNAs. We show that ribosome-, but not polysome-profiling, is biased towards identification of TOP mRNAs as differentially translated while obscuring detection of changes in non-TOP mRNA translation. Transcription start site profiling by Nano-Cap Analysis of Gene Expression (nanoCAGE) revealed that many mTOR-sensitive mRNAs do not have 5âTOP motifs. Moreover, nanoCAGE showed that 5â UTR features distinguish two functionally and translationally distinct subsets of mTOR-sensitive mRNAs: i) those with short 5â UTRs enriched for mitochondrial functions such as respiration, that are translated in an eIF4E, but not eIF4A1-dependent manner and ii) mRNAs encoding proliferation- and survival-promoting proteins, that harbor long 5â UTRs, and require both eIF4E and eIF4A1 for their efficient translation. Selective inhibition of translation of mRNAs harboring long 5â UTRs via suppression of eIF4A leads to uncoupling of expression of proteins involved in respiration (e.g. ATP5O) from those protecting mitochondrial integrity (e.g. BCL-2) ultimately resulting in apoptosis. Conversely, simultaneous translational downregulation of both long and short 5â UTR mRNAs by mTOR inhibitors results in suppression of mitochondrial respiration and predominantly cytostatic effects. Therefore, 5â UTR features define differential modes of translation of functionally distinct mTOR-sensitive mRNAs, which explains discrepancies between the effects of mTOR and eIF4A inhibitors on neoplastic cells. Determination of 5'UTR lengths using nanoCAGE in MCF7 cells
2016-03-14 | E-GEOD-77033 | biostudies-arrayexpress
Project description:Expanded CGG repeats in the 5’UTR of fragile X messenger ribonucleoprotein 1 (FMR1) gene are translated to mutant protein containing polyglycine stretches (FMRpolyG), which evoke development of neurodegenerative disorder the fragile X-associated tremor/ataxia syndrome (FXTAS). However, little is known about the regulation of noncanonical translation of mutant FMR1 mRNA. Here, we show that insulin growth factor 2 binding proteins (IGF2BPs) directly bind to 5’UTR of FMR1 mRNA to selectively enhance biosynthesis of FMRpolyG.
2025-11-21 | PXD053905 | Pride
Project description:The aim of the project is to identify RNA-binding proteins (RBPs) that interact with the 3’ untranslated region (3’UTR) of the programmed cell death 4 (PDCD4) mRNA. PDCD4 is an inflammatory tumor suppressor gene. The translation of PDCD4 mRNA is regulated by multiple RBPs. The aim is to identify RBPs which are critical for regulating PDCD4 mRNA translation by binding to its 3’UTR. For this purpose, a biotinylated PDCD4 3’UTR RNA was incubated with cytoplasmic lysate from MCF7 human breast carcinoma cells and the RNA-protein complexes pulled down by streptavidin affinity chromatography. The proteins were then eluted and resolved by SDS-PAGE. One particular protein band, migrating close to 55 KDa marker was cut out and submitted for proteomic analysis by in gel digestion and mass spectrometry.
2020-12-11 | PXD017376 | Pride
Project description:VEGF165 is one of the key regulators of tumor development. Using HCT116 cells transfected with full-length vegf mRNA, full-length vegf mRNA with mutations of H9D and L14E, mutated vegf mRNAs lacking untranslated regions (UTRs), and 5â??UTR mutated between nt 591 and nt 746, we found that vegf 5â??UTR resided an anti-apoptotic activity against chemotherapy independently of VEGF165. We next established HCT116 clones stably expressing vegf 5â??UTR or the mutated vegf 5â??UTR. The cells expressing 5â??UTR, but not the mutated UTR, showed the drug-resistant phenotype, anchorage-independent growth, and rapid tumor growth when implanted in athymic nude mice. Microarray and real-time PCR showed that the vegf 5â??UTR-expressing tumors up-regulated anti-apoptotic genes including mia and down-regulated pro-apoptotic genes including fas, pdcd1, nrg1, and bax. Microarray analysis also revealed specific down-regulation of IFN-inducible genes (43 genes) in the growing tumors. HCT116 cells stably transcribing vegf 5â??UTR decreased STAT1 expression and IFN alpha-STAT1 pathway. In addition to 5-fluorouracil treatment, the vegf 5â??UTR-expressing tumors did not respond to IFN alpha therapy at all. This novel tumor-promoting function in the vegf 5â??UTR may partly explain the insufficiency of the VEGF-VEGFR strategies and suggest a vegf-targeted silencing strategy as a more effective anti-tumor therapy. Experiment Overall Design: We established HCT116 clones stably expressing vegf 5â??UTR (vegf 5â??-G5) and vegf 5â??UTR mutated between nt 591 and nt 746 (vegf 5â?? mut-D5) and implanted into the left and the right flanks of the identical mouse, respectively. The generated tumors on day 14 after injection were removed and total RNA was extracted from each tumor using RNeasy mini (Qiagen). After removing contaminated DNA by DNaseI treatment, 10 micro grams each of RNA of vegf 5â??-G5-derived tumor from 3 individual mice was mixed and prepared for microarray analysis as sample RNA. The reference RNA of vegf 5â?? mut-D5-derived tumor was arranged in the same manner. The quality of both RNA samples was confirmed using an Agilent 2100 Bioanalyzer. Microarray analysis was then performed: briefly, 1 µg of sample RNA (vegf 5â??-G5) and reference RNA (vegf 5â?? mut-D5) were converted to cDNA with oligo dT conjugated T7 promoter primer and reverse transcriptase. These cDNAs were amplified using T7 RNA polymerase while labeling with either Cy5-CTP or Cy3-CTP (Low RNA Fluorescent Linear Amplification Kit, Agilent Technologies). 750 ng of Cy3 and Cy5-labeled cRNA were mixed and hybridized on an Agilent 22k human 1A array in a hybridization oven at 65oC for 17 h as rotating. Following hybridization and washing of a slide according to manufactureâ??s protocol, the arrays were scanned using the Agilent DNA Microarray Scanner (G2565BA) and microarray images were analyzed using Agilent Feature Extraction Software (v7.5). The final signal intensity was resulted from subtracting background signal from row spot signal. The gene expression level was presented as ratio of sample intensity against reference intensity.
2008-06-16 | E-GEOD-8888 | biostudies-arrayexpress
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Project description:The 3' untranslated region (3'UTR) constitutes a major site of post-transcriptional regulation of gene expression. Sequence elements in the 3'UTR interact with trans-acting regulators such as microRNAs that affect translation and stability. The overall aim is to use a 3'RACE cloning-sequencing stragety to identify the 3'UTRs of C. elegans transcripts and explore their heterogeneity in different developmental stages and tissues. Keywords: Transcriptome analysis For data usage terms and conditions, please refer to http://www.genome.gov/27528022 and http://www.genome.gov/Pages/Research/ENCODE/ENCODEDataReleasePolicyFinal2008.pdf Raw data files are available on our FTP site: ftp://ftp.ncbi.nlm.nih.gov/pub/geosup/Series/GSE17781 pilot study [GSM443959..GSM443964]: N2 wildtype worms staged at embryo, L1, L2, L3, L4, and adult full experiment [GSM446651..GSM446661]: N2 wildtype worms staged at embryo, L1, L2, L3, L4, dauer, and adult. Illumina Genome Analyzer sequencing of isolated clones [GSM469439] 454 sequencing of RACE clones [GSM469976]
2009-09-01 | E-GEOD-17781 | biostudies-arrayexpress
Project description:We discovered that the Saccharomyces cerevisiae lysine demethylase, Jhd2 (also known as KDM5), recruits 3'UTR processing machinery and promotes alteration of 3'UTR length in a demethylase-dependent manner. Interaction of Jhd2 with both chromatin and RNA suggests that Jhd2 affects selection of polyadenylation sites through a transcription-coupled mechanism. Wild-type yeast or yeast with JHD2 deleted were grown to mid-log phase. ChIP analysis performed for H3K4me3, Pol2, IgG.
2016-03-23 | E-GEOD-67211 | biostudies-arrayexpress