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| Status |
Public on Nov 15, 2022 |
| Title |
Structure of Escherichia coli heat shock protein Hsp15 in complex with ribosomal 50S sununits bearing peptidyl-tRNA |
| Platform organism |
Escherichia coli |
| Sample organism |
Escherichia coli BW25113 |
| Experiment type |
Non-coding RNA profiling by array
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| Summary |
In Escherichia coli, the heat shock protein 15 (Hsp15) is part of the cellular response to elevated temperature. Hsp15 interacts with peptidyl-tRNA-50S complexes that arise upon dissociation of translating 70S ribosomes, and is proposed to facilitate their rescue and recycling. A previous structure of E. coli Hsp15 in complex with peptidyl-tRNA-50S complex reported a binding site located at the central protuberance of the 50S subunit. By contrast, recent structures of RqcP, the Hsp15 homolog in Bacillus subtilis, in complex with peptidyl-tRNA-50S complexes have revealed a distinct site positioned between the anticodon-stem-loop (ASL) of the P-site tRNA and H69 of the 23S rRNA. Here we demonstrate that exposure of E. coli cells to heat shock leads to dissociation of 70S ribosomes and accumulation of 50S subunits, thus identifying a natural substrate for Hsp15 binding. Additionally, we have determined a cryo-EM reconstruction of the Hsp15-50S-peptidyl-tRNA complex isolated from heat shocked E. coli cells, revealing that Hsp15 binds to the 50S-peptidyl-tRNA complex analogously to its B. subtilis homolog RqcP. Collectively, our findings support a model where Hsp15 promotes access to the A-site for putative rescue factors to release the aberrant nascent polypeptide chain.
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| Overall design |
E. coli cell lysate and Hsp15-bound (i.e. immunopreciptated, Hsp15-IP) tRNAs used for tRNA microarrays to measure tRNA abundance - 4 biological replicates in total; 3 - Hsp15-IP and 1 - total tRNA (lysate)
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| Contributor(s) |
Safdari H, Polte C, Wilson D, Ignatova Z |
| Citation(s) |
36370110 |
| Submission date |
Oct 19, 2022 |
| Last update date |
Nov 18, 2022 |
| Contact name |
Zoya Ignatova |
| E-mail(s) |
zoya.ignatova@chemie.uni-hamburg.de
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| Organization name |
Universitaet Hamburg
|
| Street address |
Martin -Luther-King-Platz 6
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| City |
Hamburg |
| ZIP/Postal code |
20146 |
| Country |
Germany |
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| Platforms (1) |
| GPL32759 |
Microarrays of Escherichia coli encoded tRNAs |
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| Samples (4)
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| Relations |
| BioProject |
PRJNA892089 |
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