Project

Discipline

non-coding RNAs; Ribosome; Translation regulation; nucleotide analog interference

Lead
Das Ribosom, aufgebaut aus ribosomaler RNA und ribosomalen Proteinen, ist für die Proteinsynthese in allen lebenden Zellen zuständig. Da das Ribosom den Hauptangriffspunkt vieler klinisch relevanter Antibiotika darstellt, ist dessen funktionelles Verständnis von potentieller Bedeutung für die Bekämpfung von Antibiotika Resistenzen und für die Erzeugung neuer anti-mikrobieller Substanzen.
Lay summary
Molekulare Einsichten in das Ribosom und die Regulation der Proteinsynthese Inhalt und Ziel des Forschungsprojektes Ziel dieses Forschungsprojektes ist es, funktionelle Zentren des Ribosoms chemisch zu modifizieren, und dadurch molekulare Details der Proteinsynthese aufzuklären. Dabei wird eine kürzlich von uns entwickelte Methode angewendet werden, die es erlaubt, stellenspezifisch chemisch modifizierte RNA Bausteine in die ribosomale RNA einzubauen. Diese Vorgangsweise vergrössert signifikant den Pool an chemisch unterschiedlichen Gruppen, die spezifisch in das Ribosom eingebracht werden können. Regulation der Proteinsynthese ermöglicht Zellen, schnell und effizient auf geänderte Umweltbedingungen reagieren zu können. Im zweiten Hauptziel dieses Forschungsvorhabens wollen wir das Potenzial von kleinen nicht-Protein-kodierenden RNAs (ncRNAs) der Bäckerhefe untersuchen, direkt ans Ribosom zu binden und dadurch die Effizienz der Proteinsynthese zu regulieren. Wissenschaftlicher und gesellschaftlicher Kontext des Forschungsprojekts Die zentrale Rolle des Ribosoms während der Genexpression spiegelt sich auch in der Tatsache wider, dass mehr als die Hälfte aller bekannten Antibiotika die Translationsmaschinerie als Angriffspunkt haben. Molekulare Einsichten in die funktionsweisen des Ribosoms sind daher nicht nur für das bessere Verständnis der Proteinbiosynthese wesentlich, sondern eröffnen auch neue Möglichkeiten zur Bekämpfung von Antibiotika-resistenten Bakterien.

Direct link to Lay Summary

Last update: 28.11.2012

Active participation

Number	Title	Start	Funding scheme

Molecular insight into ribosome functions and translation regulation1.SUMMARYThe ribosome is the largest known RNA enzyme and is regarded as one of the most ancient catalysts in biology. The ribosome, a multifunctional RNA-protein complex, translates the genome’s message into all proteins needed for life. Its crucial role in gene expression is mirrored by the fact that the ribosome represents the main target for clinical relevant antibiotics. Therefore the knowledge of its functioning is crucial for our understanding of antibiotic resistance and for the future design of new anti-microbial compounds. In this proposal we want to address two fundamental questions: (i)How does the ribosome function at the molecular and atomic level?(ii)How is the ribosome regulated during protein biosynthesis?To gain insight into these topics we will apply an experimental tool that has been established recently in my laboratory that allows manipulating individual functional groups or even single atoms of the large subunit ribosomal RNA. This technique significantly expands the chemical repertoire of residues that can be placed at specific sites within the ribosome and thus clearly exceeds the potential of standard mutagenesis. Here we want to investigate the role of two functional centers of the large ribosomal subunit, namely the ribosomal E-site as well as the peptidyl transferase center. Both of these sites have been shown to be targeted by antibiotics thus highlight their biological relevance. By establishing a novel in vitro selection regime we further want to ‘retro-evolve’ the large ribosomal subunit RNA in order to gain insight into the evolutionary history of this ancient RNA enzyme. Pivotal for cellular life is a well-controlled and coordinated gene expression system. Given the central position of translation in gene expression it is not surprising that protein synthesis is amply targeted by regulatory molecules, mostly regulatory proteins. More recently also small non-protein-coding RNAs (ncRNAs) have been added to the list of regulatory entities (e.g. microRNAs, anti-sense RNAs). However these ncRNAs regulating translation almost exclusively target the messenger RNAs rather than the ribosome directly. In this proposal we want to test the hypothesis, that small ncRNA do exist that directly bind to the ribosome and are capable of fine-tuning its activity. A recently started genomic screen in my group for small ncRNA ribo-regulators in the yeast Saccharomyces cerevisiae revealed hundreds of potential ribosome-associated ncRNAs. With the proposed research we now want to functionally characterize some of the most promising candidates to reveal their regulatory potential.Cumulatively, the proposed research will likely expand our current knowledge of ribosome functions and more precisely will shed new light on the catalytic and regulatory repertoire of non-protein-coding RNAs in biology.