MicroRNA-21 Targets Sprouty2 and Promotes Cellular Outgrowths

Sayed, Danish; Rane, Shweta; Lypowy, Jacqueline; He, Minzhen; Chen, Ieng-Yi; Vashistha, Himanshu; Yan, Lin; Malhotra, Ashwani; Vatner, Dorothy E.; Abdellatif, Maha

doi:10.1091/mbc.e08-02-0159

Cited by 337 publications

(245 citation statements)

References 60 publications

(69 reference statements)

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“…This concept was first introduced in the context of "miRNA sponges," a term coined for transgenes containing multiple miRNA-binding sites that act as competitive inhibitors of a miRNA (35). In several cases, sponges, or related constructs, have also been shown to cause a reduction in the level of the endogenous miRNA (36,37). Ameres et al (38) recently showed that extensive complementarity between a miRNA and an artificial target can induce miRNA degradation in flies, which was also observed in HeLa cells.…”

Section: Discussionmentioning

confidence: 99%

Murine cytomegalovirus encodes a miR-27 inhibitor disguised as a target

Libri

Helwak

Miesen

et al. 2011

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

130

115

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Individual microRNAs (miRNAs) are rapidly down-regulated during conditions of cellular activation and infection, but factors mediating miRNA turnover are poorly understood. Infection of mouse cells with murine cytomegalovirus (MCMV) induces the rapid down-regulation of an antiviral cellular miRNA, miR-27. Here, we identify a transcript produced by MCMV that binds to miR-27 and mediates its degradation. UV-crosslinking and high-throughput sequencing [CRAC (UV-crosslinking and analysis of cDNA)] identified MCMV RNA segments associated with the miRNA-binding protein Argonaute 2 (Ago2). A cluster of hits mapped to a predicted miR-27-binding site in the 3′UTR of the previously uncharacterized ORF, m169. The expression kinetics of the m169 transcript correlated with degradation of miR-27 during infection, and m169 expression inhibited miR-27 functional activity in a reporter assay. siRNA knockdown of m169 demonstrated its requirement for miR-27 degradation following infection and did not affect other host miRNAs. Substitution of the miR-27-binding site in m169 to create complementarity to a different cellular miRNA, miR-24, resulted in down-regulation of only miR-24 following infection. The m169 transcript is cytoplasmic, capped, polyadenylated, and interacts with miRNA-27 through seed pairing: characteristic features of the normal messenger RNA (mRNA) targets of miRNAs. This virus-host interaction reveals a mode of miRNA regulation in which a mRNA directs the degradation of a miRNA. We speculate that RNA-mediated miRNA degradation could be a more general viral strategy for manipulating host cells.V iruses devote a large portion of their genomes to strategies for manipulating host cells and evading antiviral defense mechanisms. Numerous host miRNA-binding sites have been predicted in different viral genomes, but the validity and functional relevance of most predictions remain unclear. The best studied miRNA-virus interactions demonstrate that RNA viruses can use cellular miRNAs to regulate their life cycles; for example, the interaction between hepatitis C virus and miR-122 enhances viral replication (1), whereas the interaction between HIV-1 and miR-29 mediates its localization to P bodies (2). Direct interactions between host miRNAs and viral genes can also suppress viral gene expression and replication (3-6) (reviewed in Ref. 7). However, the factors driving the evolution of these interactions remain somewhat controversial, because they may relate to viral mechanisms for persistence and latency rather than host defense (8, 9). At the same time, the expression levels of specific miRNAs can indirectly influence infections; miRNAs are key components of the innate immune response (10, 11) and exert antiviral properties by modulating host cofactors and pathways required by viruses (12-15). We previously showed that miR-27 limits the replication capacity of murine cytomegalovirus (MCMV) but is rapidly degraded during the lytic infection (16). Actinomycin D treatment upon infection prevents miR-27 down-regulation, suggest...

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Section: Discussionmentioning

confidence: 99%

Murine cytomegalovirus encodes a miR-27 inhibitor disguised as a target

Libri

Helwak

Miesen

et al. 2011

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

130

115

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“…Among the targets, we chose Spry2 containing a miR-21 seed match in its 3 0 untranslated region (UTR), as it is known to negatively regulate Ras signaling (Cabrita and Christofori, 2008). Moreover, it has been reported that miR-21 targets Spry2 in cardiocytes and colon cancer SW480 cells (Sayed et al, 2008). We constructed a reporter plasmid (wt-UTR), driven by SV40 basal promoter, harboring the 778-nucleotide wt-3 0 UTR of Spry2 at the 3 0 position of luciferase reporter gene.…”

Section: Mir-21 Targets Spry2 For Glioma Invasionmentioning

confidence: 99%

Downregulation of Spry2 by miR-21 triggers malignancy in human gliomas

et al. 2011

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Gliomas are associated with high mortality because of their exceedingly invasive character. As these tumors acquire their invasiveness from low-grade tumors, it is very important to understand the detailed molecular mechanisms of invasion onset. Recent evidences suggest the significant role of microRNAs in tumor invasion. Thus, we hypothesized that deregulation of microRNAs may be important for the malignant progression of gliomas. We found that the aberrant expression of miR-21 is responsible for glioma invasion by disrupting the negative feedback circuit of Ras/MAPK signaling, which is mediated by Spry2. Upregulation of miR-21 was triggered by tumor microenvironmental factors such as hyaluronan and growth factors in glioma cells lacking functional phosphatase and tensin homolog (PTEN), but not harboring wild-type PTEN. Consistently with these in vitro results, Spry2 protein levels were significantly decreased in 79.7% of invasive WHO grade II-IV human glioma tissues, but not in non-invasive grade I and normal tissues. The Spry2 protein levels were not correlated with their mRNA levels, but inversely correlated with miR-21 levels. Taken together, these results suggest that the posttranscriptional regulation of Spry2 by miR-21 has an essential role on the malignant progression of human gliomas. Thus, Spry2 may be a novel therapeutic target for treating gliomas.

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“…Repression of miR by corresponding miR TS is possible, but only when the miR TS are expressed from a strong promoter at very high levels and/or the transgene contains multiple tandem miR TS (X6), which preferentially contain a bulge at position 9-12 to prevent RNA interference type endonucleolytic cleavage of the miR-miR TS duplex by Argonaute 2. 26,42,43 None of these features are characteristics of the miR-regulated AAV vectors used here. Therefore, AAV vectors with miR122 TS are obviously unable to deregulate endogenous miR122 expression levels and induce side effects.…”

Section: -δδCtmentioning

confidence: 99%

microRNA122-regulated transgene expression increases specificity of cardiac gene transfer upon intravenous delivery of AAV9 vectors

et al. 2010

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Adeno-associated virus (AAV) vectors with capsids of AAV serotype 9 enable an efficient transduction of the heart upon intravenous injection of adult mice but also transduce the liver. The aim of this study was to improve specificity of AAV9 vector-mediated cardiac gene transfer by microRNA (miR)-dependent control of transgene expression. We constructed plasmids and AAV vectors containing target sites (TSs) of liver-specific miR122, miR192 and miR148a in the 3¢ untranslated region (3¢UTR) of a luciferase expression cassette. Luciferase expression was efficiently suppressed in liver cell lines expressing high levels of the corresponding miRs, whereas luciferase expression was unaffected in cardiac myocytes. Intravenous injections of AAV9 vectors bearing three repeats of miR122 TS in the 3¢UTR of an enhanced green fluorescent expression (EGFP) expression cassette resulted in the absence of EGFP expression in the liver of adult mice, whereas the control vectors without miR TS displayed significant hepatic EGFP expression. EGFP expression levels in the heart, however, were comparable between miR122-regulated and control vectors. The liver-specific de-targeting in vivo using miR122 was even more efficient than transcriptional targeting with a cardiac cytomegalovirus (CMV)-enhanced myosin light chain (MLC) promoter. These data indicate that miR-regulated targeting is a powerful new tool to further improve cardiospecificity of AAV9 vectors.

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MicroRNA-21 Targets Sprouty2 and Promotes Cellular Outgrowths

Cited by 337 publications

References 60 publications

Murine cytomegalovirus encodes a miR-27 inhibitor disguised as a target

Murine cytomegalovirus encodes a miR-27 inhibitor disguised as a target

Downregulation of Spry2 by miR-21 triggers malignancy in human gliomas

microRNA122-regulated transgene expression increases specificity of cardiac gene transfer upon intravenous delivery of AAV9 vectors

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