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| Status |
Public on Jan 10, 2023 |
| Title |
Role of eEF2 diphthamide modification in translational fidelity |
| Organisms |
Saccharomyces cerevisiae; Cricetulus griseus |
| Experiment type |
Other
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| Summary |
Diphthamide (DPH) is a conserved amino acid modification on eukaryotic translation elongation factor eEF2. We show that loss of DPH increases -1 ribosomal frameshifting at both programmed, including in SARS-CoV-2, and non-programmed sites. Ribosome profiling of yeast and mammalian cells lacking DPH reveals increased ribosomal drop-off during elongation. The drop-off defect on the long yeast MDN1 gene was suppressed by eliminating out-of-frame stop codons. Our data reveal that loss of DPH impairs the fidelity of translocation during translation elongation resulting in increased rates of ribosomal frameshifting and premature termination at out-of-frame stop codons.
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| Overall design |
Examination of yeast and mammalian cells lacking diphthamide modification in eEF2
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| Contributor(s) |
Shin B, Ivanov IP, Kim J, Kinzy TG, Dever TE |
| Citation(s) |
37246715 |
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| Submission date |
Jun 04, 2021 |
| Last update date |
Sep 12, 2023 |
| Contact name |
Byung-Sik Shin |
| E-mail(s) |
[email protected]
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| Phone |
2404831725
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| Organization name |
NIH
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| Lab |
Protein Biosynthesis
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| Street address |
6 Center Dr, BLDG 6, RM 227
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| City |
Bethesda |
| State/province |
Maryland |
| ZIP/Postal code |
20892 |
| Country |
USA |
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| Platforms (3) |
| GPL17342 |
Illumina HiSeq 2500 (Saccharomyces cerevisiae) |
| GPL27425 |
Illumina NovaSeq 6000 (Cricetulus griseus) |
| GPL27812 |
Illumina NovaSeq 6000 (Saccharomyces cerevisiae) |
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| Samples (24)
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| GSM5359298 |
eEF2-H699N, KCl, rep2 |
| GSM5359299 |
eEF2-WT, UPF1, rep1 |
| GSM5359300 |
eEF2-WT, UPF1, rep2 |
| GSM5359301 |
eEF2-H699N, UPF1, rep1 |
| GSM5359302 |
eEF2-H699N, UPF1, rep2 |
| GSM5359303 |
eEF2-WT, upf1-del, rep1 |
| GSM5359304 |
eEF2-WT, upf1-del, rep2 |
| GSM5359305 |
eEF2-H699N, upf1-del, rep1 |
| GSM5359306 |
eEF2-H699N, upf1-del, rep2 |
| GSM5359307 |
eEF2-WT, MDN1-LS, rep1 |
| GSM5359308 |
eEF2-WT, MDN1-LS, rep2 |
| GSM5359309 |
eEF2-H699N, MDN1-LS, rep1 |
| GSM5359310 |
eEF2-H699N, MDN1-LS, rep2 |
| GSM5359311 |
CHO K1 WT, rep1 |
| GSM5359312 |
CHO K1 WT, rep2 |
| GSM5359313 |
CHO K1 dph2, rep1 |
| GSM5359314 |
CHO K1 dph2, rep2 |
| GSM5359315 |
DPH2, DTA induction, rep1 |
| GSM5359316 |
DPH2, DTA induction, rep2 |
| GSM5359317 |
dph2-del, DTA induction, rep1 |
| GSM5359318 |
dph2-del, DTA induction, rep2 |
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| Relations |
| BioProject |
PRJNA735209 |
| SRA |
SRP322739 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE176169_RAW.tar |
38.5 Mb |
(http)(custom) |
TAR (of TXT, WIG) |
SRA Run Selector |
| Raw data are available in SRA |
| Processed data provided as supplementary file |
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