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In vivo evidence that eIF3 stays bound to ribosomes elongating and terminating on short upstream ORFs to promote reinitiation
| Content Provider | Semantic Scholar |
|---|---|
| Author | Mohammad, Mahabub Pasha Pondělíčková, Vanda Munzarová Zeman, Jakub Banert Radovan Gunišová, Stanislava Valášek, Leoš Shivaya |
| Copyright Year | 2017 |
| Abstract | Translation reinitiation is a gene-specific translational control mechanism characterized by the ability of some short upstream ORFs to prevent recycling of the post-termination 40S subunit in order to resume scanning for reinitiation downstream. Its efficiency decreases with the increasing uORF length, or by the presence of secondary structures, suggesting that the time taken to translate a uORF is more critical than its length. This led to a hypothesis that some initiation factors needed for reinitiation are preserved on the 80S ribosome during early elongation. Here, using the GCN4 mRNA containing four short uORFs, we developed a novel in vivo RNA-protein Ni2+-pull down assay to demonstrate for the first time that one of these initiation factors is eIF3. eIF3 but not eIF2 preferentially associates with RNA segments encompassing two GCN4 reinitiation-permissive uORFs, uORF1 and uORF2, containing cis-acting 5΄ reinitiation-promoting elements (RPEs). We show that the preferred association of eIF3 with these uORFs is dependent on intact RPEs and the eIF3a/TIF32 subunit and sharply declines with the extended length of uORFs. Our data thus imply that eIF3 travels with early elongating ribosomes and that the RPEs interact with eIF3 in order to stabilize the mRNA-eIF3-40S post-termination complex to stimulate efficient reinitiation downstream. |
| Starting Page | 2658 |
| Ending Page | 2674 |
| Page Count | 17 |
| File Format | PDF HTM / HTML |
| Alternate Webpage(s) | https://oup.silverchair-cdn.com/oup/backfile/Content_public/Journal/nar/45/5/10.1093_nar_gkx049/1/gkx049_Supp.pdf?Expires=1494239457&Key-Pair-Id=APKAIUCZBIA4LVPAVW3Q&Signature=W8uq0Ygiew4t1x4xIf8Vw3FfshxB4sYbXtHFlyu~gKoOix8UUPbBIv52ymwm-T59gG9aGhDHGE36fRwz2OGYDgGBJ3b4VL3ljVBX9uPT1tmJyy8SGbIsW2nh2V2EWUhJN7iccGt9B1mTTtqEzbvkK0mmEsyhYCjNnXUznmItsoJBho~XbW97vjSmr-eUYOJef0OPMjcah~EsXnR6ihdqzFRIUoeJXv4GFa14NqIGaXm3q4Hi5TxDEKIAOb4ze1hGHU1SKyR5Y8rggsg0jLa-OWyZmBWqETcRrfZFygmziaB6u163Bc876BTSFlxmVKSYv2yDr~Qyzd4dSqZql1yniw__ |
| PubMed reference number | 28119417v1 |
| Alternate Webpage(s) | https://doi.org/10.1093/nar/gkx049 |
| DOI | 10.1093/nar/gkx049 |
| Journal | Nucleic acids research |
| Volume Number | 45 |
| Language | English |
| Access Restriction | Open |
| Subject Keyword | Biologic Preservation Eukaryotic Initiation Factor-3 Genes, vif Genetic Translation Process Open Reading Frames Peptide Initiation Factors Promotion (action) RNA Ribosomes Transcription Initiation Wakefulness-Promoting Agents eukaryotic translation initiation factor 3 complex location translation reinitiation |
| Content Type | Text |
| Resource Type | Article |
