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miR-196b-mediated translation regulation of mouse insulin2 via the 5'UTR
Amaresh C Panda1, Itishri Sahu2, Shardul D Kulkarni2
1National Centre for Cell Science, Ganeshkhind, Pune, India; Laboratory of Genetics, National Institute on Aging, NIH, Baltimore, Maryland, United States of America.
Abstract:
The 5' and the 3' untranslated regions (UTR) of the insulin genes are very well conserved across species. Although microRNAs (miRNAs) are known to regulate insulin secretion process, direct regulation of insulin biosynthesis by miRNA has not been reported. Here, we show that mouse microRNA miR-196b can specifically target the 5'UTR of the long insulin2 splice isoform. Using reporter assays we show that miR-196b specifically increases the translation of the reporter protein luciferase. We further show that this translation activation is abolished when Argonaute 2 levels are knocked down after transfection with an Argonaute 2-directed siRNA. Binding of miR-196b to the target sequence in insulin 5'UTR causes the removal of HuD (a 5'UTR-associated translation inhibitor), suggesting that both miR-196b and HuD bind to the same RNA element. We present data suggesting that the RNA-binding protein HuD, which represses insulin translation, is displaced by miR-196b. Together, our findings identify a mechanism of post-transcriptional regulation of insulin biosynthesis.
Insights
MicroRNA miR-196b enhances insulin biosynthesis by targeting the insulin2 gene's 5' untranslated region (UTR). This microRNA (miRNA) displaces a translation inhibitor, revealing a novel post-transcriptional regulatory mechanism for insulin production.
Area of Science:
- Molecular Biology
- Genetics
- Endocrinology
Background:
- MicroRNAs (miRNAs) regulate insulin secretion, but their direct role in insulin biosynthesis remains unclear.
- The 5' and 3' untranslated regions (UTRs) of insulin genes are highly conserved across species.
- Post-transcriptional regulation plays a crucial role in gene expression control.
Purpose of the Study:
- To investigate the direct role of microRNAs in regulating insulin biosynthesis.
- To identify specific microRNAs targeting insulin gene regulatory elements.
- To elucidate the mechanism of microRNA-mediated regulation of insulin translation.
Main Methods:
- Reporter assays to assess translation activation by miR-196b.
- Argonaute 2 knockdown experiments to confirm miRNA-dependent effects.
- RNA-binding protein (HuD) analysis to identify regulatory interactions.
Main Results:
- Mouse microRNA miR-196b specifically targets the 5'UTR of the long insulin2 splice isoform.
- miR-196b was shown to increase reporter protein translation, an effect dependent on Argonaute 2.
- miR-196b binding to the insulin 5'UTR displaces the translation inhibitor HuD.
Conclusions:
- miR-196b activates insulin translation by binding to the insulin2 5'UTR.
- This interaction leads to the displacement of the repressor protein HuD.
- A novel mechanism for post-transcriptional regulation of insulin biosynthesis involving miRNA-mediated HuD displacement is identified.
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