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Comparing proteomics and RISC immunoprecipitations to identify targets of Epstein-Barr viral miRNAs [RISC-IP-seq]

ABSTRACT: Epstein-Barr virus is a gamma-herpes virus that is causally associated with several lymphomas and carcinomas. This virus encodes at least 25 pre-miRNAs, which are expressed in infected cells to yield more than 50 detected mature miRNAs. miRNAs are small, non-coding RNAs that inhibit gene expression by promoting the inhibition of translation or of degradation of mRNAs. Currently, the function of these viral miRNAs and the contribution they provide to EBV's life-cycle remain largely unknown, due to difficulties in identifying cellular and viral genes regulated by these miRNAs. We have compared and contrasted two methods to identify targets of viral miRNAs in order to identify the advantages and limitations of each method to aid in uncovering the functions of EBV's miRNAs. Examination of RISC (RNA Induced Silencing Complexes) associated transcripts under 2 conditions in BJAB cells

ORGANISM(S): Homo sapiens

SUBMITTER: Bill Sugden

PROVIDER: E-GEOD-46514 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Comparing proteomics and RISC immunoprecipitations to identify targets of Epstein-Barr viral miRNAs [RISC-IP-seq]

Project description:Epstein-Barr virus is a gamma-herpes virus that is causally associated with several lymphomas and carcinomas. This virus encodes at least 25 pre-miRNAs, which are expressed in infected cells to yield more than 50 detected mature miRNAs. miRNAs are small, non-coding RNAs that inhibit gene expression by promoting the inhibition of translation or of degradation of mRNAs. Currently, the function of these viral miRNAs and the contribution they provide to EBV's life-cycle remain largely unknown, due to difficulties in identifying cellular and viral genes regulated by these miRNAs. We have compared and contrasted two methods to identify targets of viral miRNAs in order to identify the advantages and limitations of each method to aid in uncovering the functions of EBV's miRNAs.

2013-05-01 | GSE46514 | GEO

Epstein-Barr virus maintains lymphomas via its miRNAs

Project description:EpsteinM-bM-^@M-^PBarr virus (EBV) has evolved exquisite controls over its host cells, human B lymphocytes, not only directing these cells during latency to proliferate and thereby expand the pool of infected cells, but also to survive and thereby persist for the lifetime of the infected individual. Although these activities ensure the virus is successful, they also make the virus oncogenic, particularly when infected people are immunosuppressed. Here we show, strikingly, that one set of EBVM-bM-^@M-^Ys miRNAs both sustain BL (BurkittM-bM-^@M-^Ys lymphoma) cells in the absence of other viral oncogenes and promote the transformation of primary B lymphocytes. BurkittM-bM-^@M-^Ys Lymphoma cells were engineered to lose EBV and found to die by apoptosis and could be rescued by constitutively expressing viral miRNAs in them. Two of these EBV miRNAs were found to target Caspase 3 to inhibit apoptosis at physiological concentrations. Examination of RISC associated transcripts under 4 conditions in Sav S1-1 cells

2013-04-30 | E-GEOD-44769 | biostudies-arrayexpress

Involvement of viral miRNA in the regulation of antiviral apoptosis in shrimp

Project description:In this study, the viral miRNAs from white spot syndrome virus (WSSV) were characterized in shrimp in vivo. On the basis of our previous study and small RNA sequencing in this study, a total of 89 putative WSSV miRNAs were identified. As revealed by miRNA microarray analysis, the expressions of viral miRNAs were tissue-specific in vivo. In this study, the viral miRNAs from white spot syndrome virus (WSSV) were characterized in shrimp in vivo. On the basis of our previous study and small RNA sequencing in this study, a total of 89 putative WSSV miRNAs were identified. As revealed by miRNA microarray analysis and Northern blots, the expressions of viral miRNAs were tissue-specific in vivo. Therefore, our study presented the first report on the in vivo molecular events of viral miRNA in the antiviral apoptosis.

2014-01-09 | E-GEOD-53900 | biostudies-arrayexpress

Small RNA deep-sequencing identifies microRNAs and other small non-coding RNAs from human herpesvirus 6 (HHV-6B)

Project description:Roseolovirus, or human herpesvirus 6 (HHV-6) is a ubiquitous human pathogen infecting over 95% of the population by the age of two years. As with other herpesviruses, reactivation of HHV-6 can present with severe complications in immunocompromised individuals. Recent studies have highlighted the importance of herpesvirus-derived micro (mi)RNAs in modulating both cellular and viral gene expression. An initial report, which computed the likelihood of various viruses to encode for miRNAs, did not predict HHV-6 miRNAs. To experimentally screen for small HHV-6 encoded RNAs, we conducted large-scale sequencing of Sup-T-1 cells lytically infected with a laboratory strain of HHV-6B. This revealed an abundant 60-65 nucleotide RNA of unknown function derived from the lytic origin of replication (OriLyt) that gave rise to smaller RNA species of 18-19 nucleotides in length. In addition, we identified four pre-miRNAs, whose mature forms accumulated in Argonaute 2. In contrast to other beta-herpesviruses, HHV-6B miRNAs are expressed from direct repeat regions (DRL and DRR) located at either side of the genome. All miRNAs are conserved in the closely related HHV-6A variant, and one of them is a seed ortholog of the human miR-582-5p. Similar to alpha-herpesvirus miRNAs, they are expressed antisense to immediate early ORFs and thus have the potential to regulate key viral regulators. Small RNA sequencing from total RNA or Ago2 associated small RNAs extracted from HHV-6 infected Sup-T-1 cells

2012-01-01 | E-GEOD-34196 | biostudies-arrayexpress

Viral microRNA expression in two nasal NK T cell lymphoma cell lines

Project description:We have used microarray profiling to examine the expression of viral microRNAs (miRNAs) in two independently derived nasal natural killer T cell lymphoma (NKTCL) cell lines, SNK6 and SNT16. The goal of the study was to investigate the role of the viral miRNAs in the genesis and maintenance of NKTCLs. This experiment includes a total of 2 samples, one from SNK6 cells and the other from SNT16 cells.

2011-11-28 | E-GEOD-30695 | biostudies-arrayexpress

IGF2BP3 controls cancer cell invasiveness by modulation RISC function

Project description:The goal of this study was to determine IGF2BP3 regulation of RNA targets in human pacreatic ductal adenocarcinoma cell lines Included are iCLIP-seq libraries for IGF2BP3 from PL45 and Panc1 PDAC cell samples, RIP-seq samples from PL45 and Panc1 PDAC cells, RNA-seq data sets from control and IGF2BP3 knockdown in PL45 and Panc1 PDAC cells, and small RNA-seq samples from Panc1 cells

2016-03-12 | E-GEOD-79147 | biostudies-arrayexpress

Distinct expression pattern of miRNAs in MarekM-bM-^@M-^Ys disease virus infected-chicken splenic tumors and non-tumorous spleen tissues

Project description:To gain more insight into the molecular mechanisms of the tumorigenesis of MDV, we used microarrays to screen host and viral miRNAs that were sensitive to infection by MDV. Microarray analysis showed significant differential expression of 79 miRNAs, To determine whether miRNAs were involved in the MDV-induced tumorigenesis, miRNA microarray analysis was performed on GA-infected splenic tumors, GA-infected non-tumorous spleen tissues, and control spleen tissues at 28 dpi.

2014-04-01 | E-GEOD-56364 | biostudies-arrayexpress

Degradation of cellular miR-27 by a novel, highly abundant viral transcript is important for efficient virus replication in vivo.

Project description:Cytomegaloviruses express large amounts of viral miRNAs during lytic infection, yet, they only modestly alter the cellular miRNA profile. The most prominent alteration upon lytic murine cytomegalovirus (MCMV) infection is the rapid degradation of the cellular miR-27a and miR-27b. Here, we report that this regulation is mediated by the <1.7 kb spliced and highly abundant MCMV m169 transcript. Specificity to miR-27a/b is mediated by a single, apparently optimized, miRNA binding site located in its 3'-UTR. This site is easily and efficiently retargeted to other cellular and viral miRNAs by target site replacement. Expression of the 3'-UTR of m169 by an adenoviral vector was sufficient to mediate its function, indicating that no other viral factors are essential in this process. Degradation of miR-27a/b was found to be accompanied by 3'-tailing and -trimming. Despite its dramatic effect on miRNA stability, we found this interaction to be mutual, indicating potential regulation of m169 by miR-27a/b. Most interestingly, three mutant viruses no longer able to target miR-27a/b, either due to miRNA target site disruption or target site replacement, showed significant attenuation in multiple organs as early as 4 days post infection, indicating that degradation of miR-27a/b is important for efficient MCMV replication in vivo. Small RNA sequencing from total RNA or Ago2 associated small RNAs extracted from mock- or MCMV-infected NIH-3T3 cells

2012-03-01 | E-GEOD-34475 | biostudies-arrayexpress

Epstein-Barr Viral miRNAs Broadly Inhibit Adaptive Immunity Early in Infection

Project description:We revealed that, early in infection of human primary B cells, EBV's miRNAs regulate a wide array of immune functions encompassing antigen processing, HLAs and co-stimulatory molecules, and cytokine-cytokine receptor interactions.

2016-08-09 | GSE75776 | GEO

Comparison of DEV infected CEF with control

Project description:Duck enteritis virus (DEV) is an important herpesvirus pathogen of waterfowl associated with an acute, highly contagious lethal disease. Using a deep sequencing approach on RNA from infected chicken embryo fibroblast (CEF) cultures, we determined the global changes in the microRNA (miRNA) expression profiles during DEV infection. In addition to the changes in the expression of a number of host miRNAs as a result of DEV infection, we identified several novel DEV-encoded miRNAs. Unlike most Mardivirus-encoded miRNAs, the majority of the DEV miRNAs were encoded within the unique long region of the viral genome. The precursors of DEV miR-D18 and miR-D19 overlapped with each other suggesting similarities to miRNA-offset RNAs, although only the DEV-miR-D18-3p was functional in reporter assays. Identification of these novel miRNAs will add to the growing list of virus-encoded miRNAs enabling the exploration of their roles in pathogenesis. Each microRNA is spotted on the array 6 times. We compared expression of duck enteritis virus (DEV)-infected chicken embryo fibroblasts (CEF) with CEF control.

2012-03-08 | E-GEOD-36258 | biostudies-arrayexpress

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