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Systematic identification and characterization of RNA-binding proteins

English title Systematic identification and characterization of RNA-binding proteins
Applicant Gerber André
Number 112235
Funding scheme Project funding (Div. I-III)
Research institution Institut für Pharmazeutische Wissenschaften ETH Zürich
Institution of higher education ETH Zurich - ETHZ
Main discipline Molecular Biology
Start/End 01.05.2006 - 30.04.2009
Approved amount 197'825.00
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Keywords (6)

gene expression; eukaryotes; posttranscriptional regulation; RNA-binding protein; genomics; DNA microarrays

Lay Summary (English)

Lead
Lay summary
Post-transcriptional regulation of gene expression plays important roles in diverse cellular processes such as development, metabolism and neurogenesis. This regulation is often mediated by specific RNA-binding proteins (RBPs) that bind to cis-acting elements in the untranslated regions (UTRs) of mRNAs and regulate the stability, translation, or localization of the mRNA. Many RBPs contain characteristic RNA-binding motifs, but not every protein that binds RNA bears one of the ~100 RNA-recognition motifs that have been classified so far. To systematically identify (novel) RNA-binding proteins, we will establish an unbiased and global approach using protein microarrays. This should result in a list of novel potential RBPs. We will further select a subfraction of these to perform RNA affinity purification with subsequent DNA microarrays to identify the RNAs associated with these novel RBPs. The RNA-recognition sites of those mRNAs should be verified and mapped using in vitro assays with recombinant RBPs and synthetic RNAs, and RNA-binding should be monitored in vivo using the yeast-three hybrid assay. Finally, the regulatory functions of the selected novel RBPs should be explored by investigating their implications in translation, localization and decay of the target mRNAs and by analyzing the effect of small molecules on both the RNA-binding property and on possible other functions mediated by multifunctional proteins.
Direct link to Lay Summary Last update: 21.02.2013

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