Schematic view of the steps of ribosome biogenesis

Ribosome Assembly and Drug Screening

Ribosomes are essential macromolecular machines that translate the genetic information into proteins. To provide cells with the appropriate number of translation apparatuses the precise coordination of a large variety of synthesis and assembly steps to produce new ribosomal subunits is required. We design and apply fluorescently labeled ribosomes ("Fluorosomes") to investigate ribosomal assembly. Moreover, we use this technique to screen for potent inhibitors of bacterial ribosome biogenesis.

The figure below shows live E. coli cells with mCherry-labeled ribosomes and DAPI-stained DNA.

  

Microscope image of fluorescently labeled ribosomes in bacteria

     

Ribosome profiling via sucrose density gradient centrifugation allows to biochemically characterize ribosome biogenesis in wild type and "Fluorosome"-containing mutant cells under different conditions.

The figure below shows ribosome profiles of  E. coli cells with mCherry- and mAzami-labeled ribosomes and conditional gene knock outs that lack either the ribosomal protein S17 or L3. Strongly altered ribosome and polysome assembly is evident. For details see Nikolay et al. 2016

  

View of the data of a ribosome profiling experiment

   

Selected Publications

Nikolay, R., Hilal, T., Schmidt, S., Qin, B., Schwefel, D., Vieira-Vieira, C.H., Mielke, T., Bürger, J., Loerke, J., Amikura, K., Flügel, T., Ueda, T., Selbach, M., Deuerling, E., and Spahn, C.M.T. (2021) Snapshots of native pre-50S ribosomes reveal a biogenesis factor network and evolutionary specialization. Mol. Cell 81, 1200-1215 short | full

Nikolay, R., Schmidt, S., Schlömer, R., Deuerling, E., and Nierhaus, K. (2016) Ribosome assembly as antimicrobial target. Antibiotics 5, 18 short | full

Nikolay, R., Schlömer, R., Müller, S., and Deuerling, E. (2015) Fluorescence-based monitoring of ribosome assembly landscapes. BMC Mol. Biol. 16, 3 full

Nikolay, R., Schlömer, R., Schmidt, S., Müller, S., Heubach, A., and Deuerling, E. (2014) Validation of a fluorescence-based screening concept to identify ribosome assembly defects in Escherichia coli. Nucleic Acids Res. 42, e100 short | full