Kundu, Pradipta. Role of RNA binding protein HuR in antagonizing the microRNA-mediated repression. 2011, PhD Thesis, University of Basel, Faculty of Science.
Official URL: http://edoc.unibas.ch/diss/DissB_9709
In the present study, we have addressed these questions, using different cell-based and in vitro assays. Using mutants of HuR accumulating in the cytoplasm in the absence of stress and tumor cell lines constitutively accumulating endogenous HuR in the cytoplasm, we were able to uncouple the HuR effect on miRNA repression from stress. We also found that Ago2 and HuR do not interact with each other and that their binding to target mRNA appears to be largely mutually exclusive. Using an in vitro system with purified miRISC and recombinant HuR and its mutants, we demonstrate that HuR, by oligomerizing along RNA, leads to displacement of miRISC from RNA, even when miRISC is positioned at a distance from the primary HuR-binding site. Further, we show that HuR association with AREs can inhibit RISC-mediated endonucleolytic cleavage of target RNAs both in vivo and in vitro, and also miRNA-mediated deadenylation of RNA in the Krebs-2 ascites extract.
|Committee Members:||Muehlemann, Oliver|
|Faculties and Departments:||09 Associated Institutions > Friedrich Miescher Institut FMI|
|Bibsysno:||Link to catalogue|
|Number of Pages:||105 S.|
|Last Modified:||30 Jun 2016 10:42|
|Deposited On:||07 Feb 2012 13:21|
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