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Viewing pre-60S maturation at a minute's timescale
Gertrude Zisser1, Uli Ohmayer2, Christina Mauerhofer1
1Institute of Molecular Biosciences, Humboldtstrasse 50/EG, University of Graz, A-8010 Graz, Austria.
Nucleic Acids Research
|January 3, 2018
Summary
Diazaborine inhibits Drg1, halting ribosome biogenesis by preventing the recycling of essential proteins. This study reveals the real-time chronological order of protein assembly and RNA processing during ribosome maturation.
Area of Science:
- Molecular Biology
- Cell Biology
- Biochemistry
Background:
- Ribosome subunit formation is a complex nuclear process involving over 200 trans-acting factors.
- Some factors shuttle between the nucleus and cytoplasm, requiring efficient recycling for continued ribosome biogenesis.
Purpose of the Study:
- To investigate the real-time dynamics of ribosome maturation using a specific inhibitor.
- To establish a chronological context for protein assembly, disassembly, and pre-ribosomal RNA processing.
Main Methods:
- Utilized the inhibitor diazaborine to rapidly halt cytoplasmic release and recycling of pre-60S maturation factors.
- Tracked pre-ribosomes from nucleolar 27SA2 pre-rRNA stage to near-mature 60S subunits post-cytoplasmic export.
Main Results:
- Diazaborine inhibits the AAA-ATPase Drg1, blocking the recycling of shuttling pre-60S maturation factors.
- Inhibition leads to nucleolar depletion of these factors, arresting ribosome maturation at an early stage.
- Real-time analysis provided the first chronological framework for ribosome maturation events.
Conclusions:
- The study elucidates the temporal order of protein-RNA interactions and processing steps in ribosome biogenesis.
- Understanding these linkages is crucial for comprehending ribosome assembly defects and developing targeted interventions.