Transfer of genetic information from genes into proteins is mediated by messenger RNA (mRNA) that must be first recruited to ribosomal pre-initiation complexes (PICs) by a mechanism that is still poorly understood. Recent studies showed that besides eIF4F and poly(A)-binding protein, eIF3 also plays a critical role in this process, yet the molecular mechanism of its action is unknown. We showed previously that the PCI domain of the eIF3c/NIP1 subunit of yeast eIF3 is involved in RNA binding. To assess the role of the second PCI domain of eIF3 present in eIF3a/TIF32, we performed its mutational analysis and identified a 10-Ala-substitution (Box37) that severely reduces amounts of model mRNA in the 43-48S PICs in vivo as the major, if not the only, detectable defect. Crystal structure analysis of the a/TIF32-PCI domain at 2.65-Å resolution showed that it is required for integrity of the eIF3 core and, similarly to the c/NIP1-PCI, is capable of RNA binding. The putative RNA-binding surface defined by positively charged areas contains two Box37 residues, R363 and K364. Their substitutions with alanines severely impair the mRNA recruitment step in vivo suggesting that a/TIF32-PCI represents one of the key domains ensuring stable and efficient mRNA delivery to the PICs.
- MeSH
- alanin genetika MeSH
- eukaryotický iniciační faktor 3 chemie genetika metabolismus MeSH
- fenotyp MeSH
- iniciace translace peptidového řetězce * MeSH
- malé podjednotky ribozomu eukaryotické metabolismus MeSH
- messenger RNA metabolismus MeSH
- molekulární modely MeSH
- mutace MeSH
- Saccharomyces cerevisiae - proteiny chemie genetika metabolismus MeSH
- substituce aminokyselin MeSH
- terciární struktura proteinů MeSH
- transkripční faktory bZIP genetika MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- alanin MeSH
- eukaryotický iniciační faktor 3 MeSH
- GCN4 protein, S cerevisiae MeSH Prohlížeč
- messenger RNA MeSH
- NIP1 protein, S cerevisiae MeSH Prohlížeč
- RPG1 protein, S cerevisiae MeSH Prohlížeč
- Saccharomyces cerevisiae - proteiny MeSH
- transkripční faktory bZIP MeSH
