Systematic Analysis of Viral and Cellular MicroRNA Targets in Cells Latently Infected with Human γ-Herpesviruses by RISC Immunoprecipitation Assay

Dölken, Lars; Malterer, Georg; Erhard, Florian; Kothe, Sheila; Friedel, Caroline C.; Suffert, Guillaume; Marcinowski, Lisa; Motsch, Natalie; Barth, Stephanie; Beitzinger, Michaela; Lieber, Diana; Bailer, Susanne M.; Hoffmann, Reinhard; Ruzsics, Zsolt; Kremmer, Elisabeth; Pfeffer, Sébastien; Zimmer, Ralf; Koszinowski, Ulrich H.; Grässer, Friedrich A.; Meister, Gunter; Haas, Jürgen

doi:10.1016/j.chom.2010.03.008

Cited by 187 publications

(221 citation statements)

References 58 publications

Supporting

Mentioning

215

Contrasting

Unclassified

Order By: Relevance

“…Total RNA was prepared from 100 l of cell lysate by using an miRNeasy kit (Qiagen) following the manufacturer's instructions. Ago2 immunoprecipitation was performed as previously described (16). Briefly, 6 g of purified monoclonal hAgo2 antibody (anti-hAgo2; 11A9) or control monoclonal bromodeoxyuridine (BrdU) antibody (Abcam) was added to 5 ml of RPMI medium and incubated with 30 l of protein G-Sepharose beads (GE Healthcare) in Pierce centrifuge columns (Thermo Scientific) with constant rotation at 4°C overnight.…”

Section: Methodsmentioning

confidence: 99%

Small RNA Deep Sequencing Identifies MicroRNAs and Other Small Noncoding RNAs from Human Herpesvirus 6B

Tuddenham

Jung

Chane-Woon-Ming

et al. 2012

J Virol

Self Cite

View full text Add to dashboard Cite

Roseolovirus, or human herpesvirus 6 (HHV-6), is a ubiquitous human pathogen infecting over 95% of the population by the age of 2 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 microRNAs (miRNAs) in modulating both cellular and viral gene expression. An initial report which computed the likelihood of various viruses to encode 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-to 65-nucleotide RNA of unknown function derived from the lytic origin of replication (OriLyt) that gave rise to smaller RNA species of 18 or 19 nucleotides. In addition, we identified four pre-miRNAs whose mature forms accumulated in Argonaute 2. In contrast to the case for other betaherpesviruses, HHV-6B miRNAs are expressed from direct repeat regions (DR L and DR R ) 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 miRNA miR-582-5p. Similar to alphaherpesvirus miRNAs, they are expressed in antisense orientation relative to immediateearly open reading frames (ORFs) and thus have the potential to regulate key viral genes.

show abstract

Section: Methodsmentioning

confidence: 99%

Small RNA Deep Sequencing Identifies MicroRNAs and Other Small Noncoding RNAs from Human Herpesvirus 6B

Tuddenham

Jung

Chane-Woon-Ming

et al. 2012

J Virol

Self Cite

View full text Add to dashboard Cite

show abstract

“…The translocase of outer mitochondrial membrane 22 (TOMM22), which serves as a receptor for the Bcl-2-associated X (BAX), is targeted by miR-BART16. Antisense knockdown of TOMM22 has been shown to inhibit the association of the pro-apoptotic protein Bcl-2-associated X (BAX) with mitochondria and thus prevent BAX-induced apoptosis (Bellot et al, 2007;Dolken et al, 2010). MiR-BART15-3p can induce apoptosis partially by inhibiting an apoptosis inhibitor, BIR repeatcontaining ubiquitin-conjugating enzyme (BRUCE) (Choi et al, 2013).…”

Section: Ebv Mirnas Suppress Apoptosis Of Infected Cells To Maintain mentioning

confidence: 99%

“…EBV miR-BART1, miR-BART2, and miR-BART22 co-target interleukin-12 (IL-12), leading to decreased T cell differentiation, activation, and recognition at different stages of infection and malignant disease (Albanese et al, 2016). EBV miR-BART3-3p weakens the cytotoxic effect by inhibiting the expression of importin 7 (IPO7), which regulates T cells activation and immune tolerance (Dolken et al, 2010). MiR-BART20-5p and miR-BART8 target and alter interferon-γ (IFN-γ) secretion in EBV-positive tumors .…”

Section: Ebv-encoded Mirnas Regulate the Immune Response By Targetingmentioning

confidence: 99%

An update: Epstein-Barr virus and immune evasion via microRNA regulation

Zuo

Yue

et al. 2017

Virol. Sin.

View full text Add to dashboard Cite

Epstein-Barr virus (EBV) is an oncogenic virus that ubiquitously establishes life-long persistence in humans.To ensure its survival and maintain its B cell transformation function, EBV has developed powerful strategies to evade host immune responses. Emerging evidence has shown that microRNAs (miRNAs) are powerful regulators of the maintenance of cellular homeostasis. In this review, we summarize current progress on how EBV utilizes miRNAs for immune evasion. EBV encodes miRNAs targeting both viral and host genes involved in the immune response. The miRNAs are found in two gene clusters, and recent studies have demonstrated that lack of these clusters increases the CD4 + and CD8 + T cell response of infected cells. These reports strongly indicate that EBV miRNAs are critical for immune evasion. In addition, EBV is able to dysregulate the expression of a variety of host miRNAs, which influence multiple immune-related molecules and signaling pathways. The transport via exosomes of EBV-regulated miRNAs and viral proteins contributes to the construction and modification of the inflammatory tumor microenvironment. During EBV immune evasion, viral proteins, immune cells, chemokines, pro-inflammatory cytokines, and pro-apoptosis molecules are involved. Our increasing knowledge of the role of miRNAs in immune evasion will improve the understanding of EBV persistence and help to develop new treatments for EBV-associated cancers and other diseases.

show abstract

“…The contribution of the BART miRNAs to the development of EBV-associated cancers of epithelial origin has only begun to be explored. Several genes involved in apoptosis are potential targets of various BART miRNAs, including PUMA, Bim, and TOMM22 (17)(18)(19). Tumor suppressor genes, such as WIF1 and APC, have also been suggested as targets (20).…”

mentioning

confidence: 99%

Infection of Epstein–Barr virus in a gastric carcinoma cell line induces anchorage independence and global changes in gene expression

Marquitz

Mathur

Shair

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

Latent infection of EBV is linked to the development of multiple cancers that have distinct patterns of expression of viral proteins and microRNAs (miRNAs). In this study, we show that in vitro infection of a gastric epithelial cell line with EBV alters growth properties and induces growth in soft agar. The infected cells have high levels of expression of a large cluster of viral miRNAs, [the BamHI A rightward transcript (BART) miRNAs] and limited viral protein expression. Expression profile microarray analysis of this cell line revealed a large number of changes in cellular expression, with decreased expression of many genes. Inhibition of the traceexpressed levels of the viral oncoprotein, latent membrane protein 1, did not affect growth or alter the pattern of cellular expression. The expression changes are highly enriched for genes involved in cell motility and transformation pathways, suggesting these changes are important for the altered growth phenotype. Importantly, the transcripts decreased by microarray are significantly enriched in both experimentally and bioinformatically predicted BART miRNA targets. The absence of viral protein expression and the enrichment for viral miRNA targets in the modulated cell genes suggest that the BART miRNAs are major contributors to the transformed growth properties of the EBV-infected cells. The ability to affect cell growth through miRNA expression without viral protein expression would be a major factor in the development of cancer in individuals with functional immune systems.gastric carcinoma | oncogenesis | herpes virus

show abstract

Systematic Analysis of Viral and Cellular MicroRNA Targets in Cells Latently Infected with Human γ-Herpesviruses by RISC Immunoprecipitation Assay

Cited by 187 publications

References 58 publications

Small RNA Deep Sequencing Identifies MicroRNAs and Other Small Noncoding RNAs from Human Herpesvirus 6B

Small RNA Deep Sequencing Identifies MicroRNAs and Other Small Noncoding RNAs from Human Herpesvirus 6B

An update: Epstein-Barr virus and immune evasion via microRNA regulation

Infection of Epstein–Barr virus in a gastric carcinoma cell line induces anchorage independence and global changes in gene expression

Contact Info

Product

Resources

About