Dresden 2026 – wissenschaftliches Programm
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BP: Fachverband Biologische Physik
BP 9: Single Molecule Biophysics
BP 9.2: Vortrag
Montag, 9. März 2026, 17:15–17:30, BAR/0205
Monitoring the ribosome dynamics at the single molecule level — •Baptiste Bouhet1, Sandra Blanchet2, Charles Truong3, Olivier Namy2, and Karen Perronet1 — 1Light, Matter and Interactions lab, Gif/Yvette, France — 2Institute for Integrative Biology of the Cell, Gif/Yvette, France — 3Centre Borelli, Gif/Yvette, France
Protein synthesis is a complex multi-step process involving factors that need to interact in a coordinated manner to properly translate mRNA. As translating ribosomes cannot be synchronized over many elongation cycles, single molecule studies, mainly using total-internal-reflexion fluorescence microscopy, have been introduced to better understand translation dynamics. We decided to monitor the passage of individual, unmodified eukaryotic ribosomes from wheat germ extracts at specific fluorescent primers hybridized along mRNA. Because of the ribosome helicase activity, the double strand formed by the oligonucleotide and the mRNA is opened while the ribosome translates this region of the mRNA. Thus, the consecutive loss of fluorescence signal of two oligonucleotides allows us to measure the translation speed distribution of single ribosomes. We use this system to measure simultaneously the initiation and the elongation kinetics for linear mRNA and during -1 frameshifting, which is induced by a secondary structure on mRNA. We are also currently developing a magnetic tweezers assay to get complementary information on the opening dynamics of these structures. Thanks to its versatility, this method is a valuable tool to investigate translation machinery modifications in human diseases.
Keywords: single molecule; fluorescence microscopy; magnetic tweezers; translation; in vitro