MoLife Research Seminar presents:
Prof. Dr. Dorothee Staiger, Universität Bielefeld, Germany
Catch me if you can: Ribonomics to identify RNA-binding protein targets in Arabidopsis
A key function for RNA-binding proteins in orchestrating plant development and environmental responses is well established. However, the lack of a genome-wide view on their in vivo binding targets and binding landscapes represents a gap in understanding the mode of action of plant RNA-binding proteins. Here, we adapt individual nucleotide resolution crosslinking immunoprecipitation (iCLIP) for genome-wide determining the binding repertoire of the circadian clock-regulated Arabidopsis thaliana glycine-rich RNA-binding protein AtGRP7. iCLIP identified transcripts with significantly enriched crosslink sites in plants expressing AtGRP7-GFP and absent in plants expressing an RNA-binding-dead AtGRP7 variant or GFP alone. AtGRP7 can bind to all transcript regions, the 3´and 5´untranslated regions, introns and coding sequences with a preference for untranslated regions. Cross-referencing the targets against transcriptome changes in AtGRP7 loss-of-function mutants or AtGRP7-overexpressing plants revealed a predominantly negative effect of AtGRP7 on its targets. In particular, elevated AtGRP7 levels lead to damping of circadian oscillations of transcripts Furthermore, several targets show changes in alternative splicing in response to altered AtGRP7 levels. Establishing iCLIP for plants to identify target transcripts of the RNA-binding protein AtGRP7 paves the way to investigate the dynamics of posttranscriptional networks in response to exogenous and endogenous cues.
All are welcome!
Further information: Prof. Dr. Christian Hammann, Professor of Biochemistry - Life Sciences & Chemistry - Email: c.hammann [at] jacobs-university.de - Tel: +49 421 200-3247 - Link to Homepage: https://www.jacobs-university.de/ses/chammann/ribogenetics