PROJECT INFORMATION :
We here propose to use mass spectrometry to identify protein binding partners at the level of individual RNA folds and to define an RBP interactome network based on quantitative interactome data. We will use our streamlined RNA-protein interactomics approach to uncover RNA-binding proteins at phylogenetic conserved mRNA folds and characterize RNA-dependent mRBP composition in S. cerevisiae. Besides uncovering systematically the functionality of these RNA readers in yeast for a systems perspective, our technique is scalable and widely applicable to any RNA structure and species. It thus can contribute to deciphering the mRNP code also in other model systems.
KEY TECHNOLOGIES :
- Mass spectrometry
- Computational analysis
Scheibe M., Arnoult N., Kappei D., Buchholz F., Decottignies A., Butter F.# and Mann M.# Quantitative interaction screen of telomeric repeat-containing RNA reveals novel TERRA regulators. Genome Res. (2013) 23, 2149-2157.
Klaas D., Scheibe M., Butter F., Hogan G.J., Mann M. and Brown P.O. Quantitative proteomic analysis reveals concurrent RNA-protein interactions and identifies new RNA-binding proteins in Saccharomyces cerevisiae. Genome Res. (2013) 23, 1028-38.
Scheibe M., Butter F., Hafner M., Tuschl T. and Mann M. Quantitative mass spectrometry and PAR-CLIP to identify RNA-protein interactions. Nucleic Acids Res. (2012) 40, 9897-902.
Butter F., Scheibe M., Mörl M. and Mann M. Unbiased RNA-protein interaction screen by quantitative proteomics. Proc. Natl. Acad. Sci. U S A (2009) 106, 10626-10631