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| Status |
Public on Aug 30, 2018 |
| Title |
Tma64 (eIF2D), Tma20 (MCT-1), and Tma22 (DENR) recycle post-termination 40S subunits in vivo |
| Organism |
Saccharomyces cerevisiae |
| Experiment type |
Other
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| Summary |
The recycling of ribosomal subunits after translation termination is critical for efficient gene expression. Tma64 (eIF2D), Tma20 (MCT-1), and Tma22 (DENR) function as 40S recycling factors in vitro, but it is unknown whether they perform this function in vivo or serve as alternative initiation factors. Ribosome profiling of strains missing these factors revealed 80S ribosomes queued behind the stop codon, consistent with a block in 40S recycling. We found that unrecycled ribosomes could reinitiate translation at AUG codons in the 3’UTR, as evidenced by peaks in the footprint data and 3’UTR reporter analysis. In vitro translation experiments using reporter mRNAs containing upstream ORFs (uORFs) further established that reinitiation increased in the absence of these proteins. In some cases, 40S ribosomes appeared to rejoin with 60S subunits and undergo an alternative 80S reinitiation process in 3’UTRs. These results support a crucial role for Tma64, Tma20, and Tma22 in the recycling of 40S ribosomal subunits at stop codons and translation reinitiation.
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| Overall design |
14 biological samples are included in the study for ribosome footprinting. These include wild-type and combination knockouts of TMA64, TMA20, and TMA22, and mutation of SUI1.
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| Contributor(s) |
Young DJ, Makeeva DS, Zhang F, Anisimova AS, Stolboushkina EA, Ghobakhlou A, Shatsky IN, Dmitriev SE, Hinnebusch AG, Guydosh NR |
| Citation(s) |
30146315 |
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| Submission date |
Jan 09, 2018 |
| Last update date |
Mar 13, 2019 |
| Contact name |
Nicholas R Guydosh |
| E-mail(s) |
[email protected]
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| Organization name |
National Institutes of Health
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| Department |
NIDDK
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| Lab |
LBG
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| Street address |
8 Center Dr, Bldg 8, 220
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| City |
Bethesda |
| State/province |
MD |
| ZIP/Postal code |
20892 |
| Country |
USA |
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| Platforms (2) |
| GPL13821 |
Illumina HiSeq 2000 (Saccharomyces cerevisiae) |
| GPL17342 |
Illumina HiSeq 2500 (Saccharomyces cerevisiae) |
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| Samples (14)
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| Relations |
| BioProject |
PRJNA429210 |
| SRA |
SRP128535 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE108942_RAW.tar |
124.0 Mb |
(http)(custom) |
TAR (of WIG) |
SRA Run Selector |
| Raw data are available in SRA |
| Processed data provided as supplementary file |
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