Prof. Dr. Sträßer
PROJECT INFORMATION :
The Hermes of gene expression is the messenger RNA (mRNA), which carries the information of a protein-coding gene to the ribosome. During nuclear mRNP biogenesis many proteins bind to the mRNA, processing and packaging it into a messenger ribonucleoprotein particle (mRNP). These RNA-binding proteins (RBPs) of the mRNP not only function at the time of their binding, but often also affect later steps of gene expression. Consequently, the process of mRNP packing in the nucleus is essential and at the same time dauntingly complex.
In the project proposed here, we combine biochemical and mass-spectrometric approaches to unravel the process of nuclear mRNP packaging and the functions of the RBPs involved. We will systematically identify the RNA-binding sites in these RBPs in S. cerevisiae by photo- and chemical cross-linking combined with high-resolution mass spectrometry. We will develop new techniques for chemical protein–RNA cross-linking that will improve mRNP analysis in the future. Using mutants in the RNA-binding sites thus identified, we will assess each RBP's function in mRNP packaging, nuclear mRNA export and mRNA stability. We expect that the results will elucidate the process of nuclear mRNP packaging and the influence of mRNP composition on the life cycle of mRNA in the cell.
KEY TECHNOLOGIES :
- Chromatin immunoprecipitation (ChIP)
- Tandem affinity purification (TAP)
- Yeast (S.c.) genetics
Reuter, M., Meinel, D.M., and Sträßer, K. (2015) The poly(A)-binding protein Nab2 functions in RNA polymerase III transcription. Genes Dev. 29: 1565-1575
Meinel, D.M. and Sträßer, K. (2015) Co-transcriptional mRNP formation is coordinated within a molecular mRNP packaging station in S. cerevisiae. BioEssays 37: 666-677 (invited review)
Meinel, D.M., Burkert-Kautzsch, C., Kieser, A., O’Duibhir, E., Siebert, M., Mayer, A., Cramer, P., Söding, J., Holstege, F.C., and Sträßer, K. (2013) Recruitment of TREX to the Transcription Machinery by its Direct Binding to the Phospho-CTD of RNA Polymerase II. PLoS Genetics 9: e1003914
Chanarat, S., Seizl, M., and Sträßer, K. (2011) The Prp19 Complex is a Novel Transcription Elongation Factor Required for TREX Occupancy at Transcribed Genes. Genes Dev. 25: 1147-1158
Sträßer, K. , Masuda, S., Mason, P., Pfannstiel J., Oppizzi M., Rodriguez-Navarro S., Rondón A.G., Aguilera A., Struhl, K., Reed R., and Hurt E. (2002) TREX is a conserved complex coupling transcription with messenger RNA export. Nature 417: 304-308
INFO EXTRA :