The interplay of HuR and miR-3134 in regulation of AU rich transcriptome
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Title |
The interplay of HuR and miR-3134 in regulation of AU rich transcriptome
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Creator |
Sharma, Shivani
Verma, Suneer Vasudevan, Madavan Samanta, Subhasis Thakur, Jitendra K. Kulshreshtha, Ritu |
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Subject |
MicroRNA
miR-3134 AU rich elements HuR Breast cancer |
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Description |
Accepted date: 20 Jun 2013
MicroRNAs and AU Rich element (ARE)-mediated degradation of transcripts are thought to be two independent means of gene regulation at the post-transcriptional level. However, since their site of action is the same (3'UTR of mRNA), there exists a high probability that specific miRNAs may bind to AREs and, thus, interact with ARE-binding proteins (ARE-BPs) to regulate transcript levels. In this study, we have characterized AREs as potential targets of hsa-miR-3134. An analysis of the global gene expression profile of breast cancer cell line MCF7 overexpressing miR-3134 revealed the presence of at least one AUUUA element in the 3'-UTRs of 63% of miR-3134 regulated protein coding genes. Quantitative RT-PCR or 3'UTR luciferase assays show that miR-3134 mediates an up to 4-8-fold increase in the levels of ARE bearing transcripts-SOX9, VEGFA, and EGFR, while mutated miR-3134 shows a decreased effect. The miR-3134-mediated increase in transcript levels was unaffected by treatment with transcription inhibitor (actinomycin D), indicating that miR-3134 enhances transcript stability. To investigate a possible interplay between miR-3134 and a prototype ARE-BP, HuR, we compared their overexpression transcriptome profiles. Interestingly, up to 80% of miR-3134-regulated genes were also regulated by HuR. Overexpression studies of HuR alone or in combination with miR-3134 shows that wt miR-3134 but not a mutated miR-3134 promotes stabilization of HuR-regulated transcripts SOX9, VEGFA, and EGFR as confirmed by qRT-PCR or RNA-immunoprecipitation experiments. Overall, this report suggests that collaboration between ARE-binding microRNAs and ARE-binding proteins could be a general mechanism of 3'-UTR mediated regulation of gene expression in human cells. RK acknowledges support from the Department of Biotechnology, GOI for a RGYI grant. We acknowledge Prof Alok Bhattacharya, School of Life Sciences, Jawaharlal Nehru University for extend- ing the lab facilities funded and supported by the Department of Biotechnology for experimental work. SS1 thanks the Department of Biotechnology for a Junior Research Fellowship. SS2 acknowledges fellowship from the DBT-RA program. JKT acknowledges funding from NIPGR core grant. We acknowledge Dr Stefan Oehler for carefully editing this manuscript. |
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Date |
2015-11-18T08:40:38Z
2015-11-18T08:40:38Z 2013 |
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Type |
Article
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Identifier |
RNA Biol., 10(8): 1283-1290
2161-5063 http://172.16.0.77:8080/jspui/handle/123456789/367 http://www.tandfonline.com/doi/full/10.4161/rna.25482 10.4161/rna.25482 |
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Language |
en_US
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Publisher |
Taylor & Francis Group
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