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Ribosome dynamics during spontaneous frameshifting

dc.contributor.advisorRodnina, Marina Prof. Dr.
dc.contributor.authorPoulis, Panagiotis
dc.date.accessioned2022-10-18T11:55:56Z
dc.date.available2022-10-25T00:50:08Z
dc.date.issued2022-10-18
dc.identifier.urihttp://resolver.sub.uni-goettingen.de/purl?ediss-11858/14295
dc.identifier.urihttp://dx.doi.org/10.53846/goediss-9493
dc.language.isoengde
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/
dc.subject.ddc570de
dc.titleRibosome dynamics during spontaneous frameshiftingde
dc.typedoctoralThesisde
dc.contributor.refereeRodnina, Marina Prof. Dr.
dc.date.examination2022-10-04de
dc.description.abstractengDuring protein synthesis, the ribosome moves along the mRNA in steps of three nucleotides at a time to maintain the translational reading frame. However, on some mRNA sequences that are denoted as slippery, ribosomes may shift by one or several nucleotides to the 5’ direction in a process called spontaneous –1 frameshifting. The result is the production of aberrant peptides with a different length and amino acid sequence than the 0-frame peptide. Frameshifting is a major potential source of translation errors, which compromise the fitness of the cell. It is thus important to understand the molecular mechanism that leads to spontaneous frameshifting. In this study, we use single molecule Förster resonance energy transfer (smFRET) to investigate how the tRNAs move through the ribosome during translocation on slippery mRNA. We investigate the internal motions of the ribosome (i.e. ribosome dynamics) and the role of EF-G during translocation on slippery mRNA. Our results show that uncoupling the movements of the two tRNA that are displaced during translocation is a key determinant of spontaneous frameshifting. While the deacylated tRNA moves rapidly from the P to the E site and is released from the ribosome, the translocation of the peptidyl-tRNA from the A to the P site is slow and stalled between intermediate (chimeric, CHI) and posttranslocation states. Delayed translocation in a different stage of translocation, i.e. prior to the formation of chimeric states, does not contribute to frameshifting. During slow translocation of the peptidyl-tRNA, the SSU head domain is in the swiveled conformation, further facilitating the positional uncertainty of the peptidyl-tRNA. This allows sampling of the 0- and –1-frame codons, thus shifting the reading frame. We anticipate our work to be the starting point for further investigation of spontaneous frameshifting, the effect of frameshifted peptides in the cell in physiological and pathological conditions, the respective mechanism of frameshifting in eukaryotes, as well as the emergence of spontaneous frameshifting as a therapeutic target in human disease and infection.de
dc.contributor.coRefereeEnderlein, Jörg Prof. Dr.
dc.subject.engribosome, smFRET, translocation, frameshiftingde
dc.identifier.urnurn:nbn:de:gbv:7-ediss-14295-0
dc.affiliation.instituteBiologische Fakultät für Biologie und Psychologiede
dc.subject.gokfullBiologie (PPN619462639)de
dc.description.embargoed2022-10-25de
dc.identifier.ppn1819406970
dc.notes.confirmationsentConfirmation sent 2022-10-18T12:15:01de


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