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| Status |
Public on Aug 18, 2021 |
| Title |
The RNA helicase Dbp7 promotes domain V/VI compaction and stabilization of inter-domain interactions during early 60S assembly |
| Organism |
Saccharomyces cerevisiae |
| Experiment type |
Other
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| Summary |
Early pre-60S ribosomal particles are poorly characterized, highly dynamic complexes that undergo extensive rRNA folding and compaction concomitant with assembly of ribosomal proteins and exchange of assembly factors. Pre-60S particles contain numerous RNA helicases, which are likely regulators of accurate and efficient formation of appropriate rRNA structures. Here we reveal binding of the RNA helicase Dbp7 to domain V/VI of early pre-60S particles in yeast and show that in the absence of this protein, dissociation of the Npa1 scaffolding complex, release of the snR190 folding chaperone, recruitment of the A3 cluster factors and binding of the ribosomal protein uL3 are impaired. uL3 is critical for formation of the peptidyltransferase center and is responsible for stabilizing interactions between the 5’ and 3’ ends of the 25S, an essential pre-requisite for subsequent pre-60S maturation events. Highlighting the importance of pre-ribosome remodeling by Dbp7, our data suggest that in the absence of Dbp7 or its catalytic activity, early pre-ribosomal particles are targeted for degradation.
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| Overall design |
Wild-type BY4741 yeast cells or cells expressing genomically His-Tev-ProtA-tagged Dbp7 were grown in the presence of 4-thiouracil, crosslinked using light at 365 nm and analysed by CRAC.
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| Contributor(s) |
Aquino GR, Hackert P, Krogh N, Pan K, Jaafar M, Henras AK, Nielsen H, Urlaub H, Bohnsack KE, Bohnsack MT |
| Citation(s) |
34686661 |
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| Submission date |
Nov 03, 2020 |
| Last update date |
Nov 17, 2021 |
| Contact name |
Katherine Bohnsack |
| E-mail(s) |
[email protected]
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| Organization name |
University Medical Centre Göttingen
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| Street address |
Humboldtallee 23
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| City |
Göttingen |
| ZIP/Postal code |
37073 |
| Country |
Germany |
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| Platforms (1) |
| GPL13821 |
Illumina HiSeq 2000 (Saccharomyces cerevisiae) |
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| Samples (4)
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| Relations |
| BioProject |
PRJNA674317 |
| SRA |
SRP290971 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE160734_RAW.tar |
150.0 Kb |
(http)(custom) |
TAR (of TXT) |
SRA Run Selector |
| Raw data are available in SRA |
| Processed data provided as supplementary file |
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