a 3′-untranslated region (3′utr) induces organ adhesion by regulating mir-199a functions3u2032未翻译区(3u2032utr)诱导器官粘连通过调节mir - 199 a的功能.pdfVIP

A 39-Untranslated Region (39UTR) Induces Organ Adhesion by Regulating miR-199a* Functions 1,2 1,2 1,2 1 1,2 Daniel Y. Lee , Tatiana Shatseva , Zina Jeyapalan , William W. Du , Zhaoqun Deng , Burton B. Yang1,2* 1 Sunnybrook Research Institute, Sunnybrook Health Sciences Centre, Toronto, Canada, 2 Department of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, Canada Abstract Mature microRNAs (miRNAs) are single-stranded RNAs of 18–24 nucleotides that repress post-transcriptional gene expression. However, it is unknown whether the functions of mature miRNAs can be regulated. Here we report that expression of versican 39UTR induces organ adhesion in transgenic mice by modulating miR-199a* activities. The study was initiated by the hypothesis that the non-coding 39UTR plays a role in the regulation of miRNA function. Transgenic mice expressing a construct harboring the 39UTR of versican exhibits the adhesion of organs. Computational analysis indicated that a large number of microRNAs could bind to this fragment potentially including miR-199a*. Expression of versican and fibronectin, two targets of miR-199a*, are up-regulated in transgenic mice, suggesting that the 3 9UTR binds and modulates miR-199a* activities, freeing mRNAs of versican and fibronectin from being repressed by miR-199a*. Confirmation of the binding was performed by PCR using mature miR-199a* as a primer and the targeting was performed by luciferase assays. Enhanced adhesion by expression of the 39UTR was confirmed by in vitro assays. Our results demonstrated that upon arrival in cytoplasm, miRNA activities can be modulated locally by the 39UTR. Our assay ma