rRNA; ribosome; non-coding RNAs; translation; RNA biology; translation regulation; nucleotide analog interference
Fricker Roger, Brogli Rebecca, Luidalepp Hannes, Wyss Leander, Fasnacht Michel, Joss Oliver, Zywicki Marek, Helm Mark, Schneider André, Cristodero Marina, Polacek Norbert (2019), A tRNA half modulates translation as stress response in Trypanosoma brucei, in Nature Communications
, 10(1), 118-118.
Shikha Shikha, Brogli Rebecca, Schneider André, Polacek Norbert (2019), tRNA Biology in Trypanosomes, in CHIMIA International Journal for Chemistry
, 73(5), 395-405.
HeissenbergerClemens, LiendlLisa, NagelreiterFabian, GonskikhYulia, YangGuohuan, StelzerElena, KrammerTeresa, MicutkovaLucia, VogtStefan, KreilDavid, SekotGerhard, SienaEmilio, PoserIna, HarreitherEva, LinderAngela, EhretViktoria, HelbichThomas, Grillari-VoglauerRegina, Jansen-DürrPidder, KosMartin, PolacekNorbert, GrillariJohannes, SchossererMarkus (2019), Loss of the ribosomal RNA methyltransferase NSUN5 impairs global protein synthesis and normal growth, in Nucleic Acids Research
Wyss Leander, Waser Melanie, Gebetsberger Jennifer, Zywicki Marek, Polacek Norbert (2018), mRNA-specific translation regulation by a ribosome-associated ncRNA in Haloferax volcanii, in Scientific Reports
, 8(1), 12502-12502.
Willi Jessica, Küpfer Pascal, Evéquoz Damien, Fernandez Guillermo, Katz Assaf, Leumann Christian, Polacek Norbert (2018), Oxidative stress damages rRNA inside the ribosome and differentially affects the catalytic center, in Nucleic Acids Research
, 46(4), 1945-1957.
Hoernes Thomas, Clementi Nina, Juen Michael, Shi Xinying, Faserl Klaus, Willi Jessica, Gasser Catherina, Kreutz Christoph, Joseph Simpson, Lindner Herbert, Hüttenhofer Alexander, Erlacher Matthias (2018), Atomic mutagenesis of stop codon nucleotides reveals the chemical prerequisites for release factor-mediated peptide release, in Proceedings of the National Academy of Sciences
Koch Miriam, Willi Jessica, Pradère Ugo, Hall Jonathan, Polacek Norbert (2017), Critical 23S rRNA interactions for macrolide-dependent ribosome stalling on the ErmCL nascent peptide chain, in Nucleic Acids Research
J Gebetsberger, L Wyss, AM Mleczko, J Reuther, N Polacek (2016), A tRNA-derived fragment competes with mRNA for ribosome binding and regulates translation during stress., in RNA biology
1.SUMMARYThe ribosome is the largest known RNA catalyst and is certainly one of the most central enzymes for cellular metabolism. Recent advances in the research field of protein biosynthesis suggest that the translation machinery serves as pivotal regulatory hub in protein homeostasis and stress response. While initially it was assumed that every ribosome in a cell or organism is identical in respect to its molecular composition and catalytic performance, a growing body of evidence argues against this view. It has been shown that translation in general and the ribosome in particular has to rapidly adapt to external signals. Especially during stress response the rate of protein biosynthesis has to be tightly adjusted in order not to waste precious cellular energy and to allow stress-response adaptation networks to be established. Failure of the translation apparatus to adequately respond to challenging environmental condition has been amply implied in cell death and disease. In this proposal we want to address two fundamental questions: (i)What are the consequences of oxidative stress on the structure and function of the ribosome?(ii)How do small ribosome-associated non-coding RNAs regulate translation?To gain insight into the consequences of oxidative stress on the ribosome, we will apply an experimental tool called “atomic mutagenesis” which allows the site specific incorporation of individual oxidized nucleobases into the ribosome. In the course of this project we will replace all inner shell active site residues by their respective oxidized nucleobase versions. By the planned experiments we will be able to gain molecular insight into the functional consequences of oxidative stress in the catalytic heart ribosome and how this might relate to disease.Accumulating recent evidence identified the ribosome as binding target for numerous small non-protein-coding RNAs (ncRNAs) in various organisms of all three domains of life. Therefore it appears that ribosome-associated ncRNAs (rancRNAs) are a prevalent, yet poorly understood class of cellular transcripts. Here we aim at elucidating the functional mechanisms how stress-induced rancRNAs are capable of fine tuning the rate of protein synthesis in a eukaryal and an archaeal model organism. A comprehensive set of in vivo and in vitro approaches will be employed to characterize the regulatory potential of various rancRNAs during a variety of cellular stress situations.Cumulatively, the proposed research will likely expand our current knowledge of how cells, and in particular how the ribosome, adapt to stress. Specifically this project will reveal the so far elusive RNA biology of the emerging class of rancRNAs during translation control.