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| Status |
Public on Apr 01, 2017 |
| Title |
Structural transitions during yeast large ribosomal subunit maturation analyzed by tethered nuclease structure probing |
| Organism |
Saccharomyces cerevisiae |
| Experiment type |
Expression profiling by high throughput sequencing
Other
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| Summary |
Yeast large ribosomal subunit (LSU) precursors are subject to substantial changes in protein composition during their maturation due to coordinated transient interactions with a large number of ribosome biogenesis factors and due to the assembly of ribosomal proteins. These compositional changes go along with stepwise processing of LSU rRNA precursors and with specific rRNA folding events, as revealed by recent cryo-electron microscopy analyses of late nuclear and cytoplasmic LSU precursors. Here we aimed to analyze changes in the spatial rRNA surrounding of selected ribosomal proteins during yeast LSU maturation. For this we combined a recently developed tethered tertiary structure probing approach with both targeted and high-throughput readout strategies. Several structural features of late LSU precursors were faithfully detected by this procedure. In addition, the obtained data let us suggest that early rRNA precursor processing events are accompanied by a global transition from a flexible to a spatially restricted rRNA conformation. For intermediate LSU precursors, a number of structural hallmarks could be addressed which include the fold of the internal transcribed spacer between 5.8S rRNA and 25S rRNA, the orientation of the central protuberance and the spatial organization of the interface between LSU rRNA domains I and III.
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| Overall design |
Major rRNA cleavages by nuclease fusion proteins incorporated in LSU precursors were identified by random primer extension reactions followed by high-throughput sequencing of the resulting cDNA fragments.
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| Contributor(s) |
Pöll G, Müller C, Bodden M, Teubl F, Eichner N, Lehmann G, Griesenbeck J, Tschochner H, Milkereit P |
| Citation(s) |
28686620 |
| |
| Submission date |
Mar 31, 2017 |
| Last update date |
May 15, 2019 |
| Contact name |
Philipp Milkereit |
| E-mail(s) |
philipp.milkereit@vkl.uni-regensburg.de
|
| Phone |
++49 941 943 2846
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| Organization name |
Universität Regensburg
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| Department |
Lehrstuhl für Biochemie III
|
| Street address |
Universitätsstr. 31
|
| City |
Regensburg |
| ZIP/Postal code |
93053 |
| Country |
Germany |
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| Platforms (1) |
| GPL17143 |
Illumina MiSeq (Saccharomyces cerevisiae) |
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| Samples (8)
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| Relations |
| BioProject |
PRJNA381162 |
| SRA |
SRP102782 |
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