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| Status |
Public on Dec 22, 2021 |
| Title |
Phosphorylation of Ribosomal Protein S6 differentially affects mRNA translation based on ORF length |
| Organisms |
Homo sapiens; Mus musculus |
| Experiment type |
Expression profiling by high throughput sequencing
|
| Summary |
Phosphorylation of Ribosomal Protein S6 (RPS6) was the first post-translational modification of the ribosome to be identified and is a commonly-used readout for mTORC1 activity. Although the cellular and organismal functions of RPS6 phosphorylation are known, its molecular consequences on translation are less well understood. Here we use selective ribosome footprinting to analyze the location of ribosomes containing phosphorylated RPS6 on endogenous mRNAs in cells. We find that RPS6 becomes progressively dephosphorylated on ribosomes as they translate an mRNA. As a consequence, average RPS6 phosphorylation is higher on mRNAs with short coding sequences (CDSs) compared to mRNAs with long CDSs. Loss of RPS6 phosphorylation causes a correspondingly larger drop in translation efficiency of mRNAs with short CDSs than long CDSs. Interestingly, mRNAs with 5’ TOP motifs are translated well also in the absence of RPS6 phosphorylation despite short CDS lengths, suggesting they are translated via a different mode. In sum this provides a dynamic view of RPS6 phosphorylation on ribosomes as they translate mRNAs and the functional consequence on translation.
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| Overall design |
We develop modification selective ribosome footprinting to determine the rate of RPS6 phosphorylation of translating ribosomes across the translatome. This allows us to gain mechanistic understanding of the role of RPS6 phosphorylation in ribosome function.
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| Contributor(s) |
Bohlen J, Teleman A, Roiuk M |
| Citation(s) |
34871442, 35869369 |
| Submission date |
Mar 16, 2021 |
| Last update date |
Jul 28, 2022 |
| Contact name |
Jonathan Bohlen |
| Organization name |
DKFZ
|
| Department |
B140
|
| Lab |
AG Teleman
|
| Street address |
Im Neuenheimer Feld 581
|
| City |
Heidelberg |
| ZIP/Postal code |
69123 |
| Country |
Germany |
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| Platforms (2) |
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| Samples (7)
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| GSM5173412 |
p-RPS6 selective Riosome footprints from HeLa cells |
| GSM5173413 |
Total ribosome fooprints from HeLa cells |
| GSM5173414 |
p-RPS6 selective Riosome footprints from WT MEF cells |
| GSM5173415 |
Total ribosome fooprints from WT MEF cells |
| GSM5173416 |
Total ribosome fooprints from p-RPS6 -/- MEF cells |
| GSM5173417 |
Total mRNA sequencing from WT MEF cells |
| GSM5173418 |
Total mRNA sequencing from p-RPS6 -/- MEF cells |
|
| Relations |
| BioProject |
PRJNA714824 |
| SRA |
SRP310845 |
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