Transcriptome-wide functional analysis of post-transcriptional regulatory interactions of the RNA-binding protein HuR/ELAVL1
Year: 2012
Type: dissertation
Abstract: Post-transcriptional regulation is performed by small RNAs and RNA-binding proteins (RBPs) which control mRNA splicing, export, degradation and translation efficiency. Each small RNA and RBP interacts with up to thousands of target transcripts, but only for a few RBPs their targets have been comprehensively identified. HuR/ELAVL1 is a conserved RBP which regulates mRNA stability and translation. The aim of the present collaborative work was to characterize functional targets of HuR. To obtain precise transcriptome-wide binding sites of HuR in human cells, we used crosslinking and immunoprecipitation (IP) coupled with deep sequencing of bound RNA. The functionality of these sites was validated by measuring the transcriptome-wide impact of HuR knock down on mRNA levels. We also measured changes in protein synthesis of thousands of proteins which reflected the role of HuR in protein synthesis. We found multiple binding sites of HuR in introns and shown a role of HuR in mRNA processing. We also discovered the unexpected regulation of the microRNA miR-7 by HuR. In summary, we identified thousands of direct and functional HuR targets, found a human miRNA controlled by HuR, and propose a role for HuR in splicing.
Author: Lebedeva, Svetlana
Language: English
Topic: RNA Research and Splicing
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: green
OpenAlex ID: W1830075815