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Updated: Jul 16, 2025

Monitoring eIF4F Assembly by Measuring eIF4E-eIF4G Interaction in Live Cells
Published on: May 1, 2020
Yeast Tor complex 1 phosphorylates eIF4E-binding protein, Caf20
Yoshiaki Kamada1,2, Ryoko Ando3, Shingo Izawa3
1National Institute for Basic Biology, Okazaki, Japan.
Abstract:
Tor complex 1 (TORC1), a master regulator of cell growth, is an evolutionarily conserved protein kinase within eukaryotic organisms. To control cell growth, TORC1 governs translational processes by phosphorylating its substrate proteins in response to cellular nutritional cues. Mammalian TORC1 (mTORC1) assumes the responsibility of phosphorylating the eukaryotic translation initiation factor 4E (eIF4E)-binding protein 1 (4E-BP1) to regulate its interaction with eIF4E. The budding yeast Saccharomyces cerevisiae possesses a pair of 4E-BP genes, CAF20 and EAP1. However, the extent to which the TORC1-4E-BP axis regulates translational initiation in yeast remains uncertain. In this study, we demonstrated the influence of TORC1 on the phosphorylation status of Caf20 in vivo, as well as the direct phosphorylation of Caf20 by TORC1 in vitro. Furthermore, we found the TORC1-dependent recruitment of Caf20 to the 80S ribosome. Consequently, our study proposes a plausible involvement of yeast's 4E-BP in the efficacy of translation initiation, an aspect under the control of TORC1.
Insights
This study shows that Tor complex 1 (TORC1) directly phosphorylates yeast
Area of Science:
- Cell Biology
- Molecular Biology
- Biochemistry
Background:
- Tor complex 1 (TORC1) regulates cell growth by controlling translation.
- In mammals, mTORC1 phosphorylates 4E-BP1, affecting translation initiation.
- The role of yeast 4E-BPs (Caf20, Eap1) in TORC1-mediated translation is unclear.
Purpose of the Study:
- To investigate the regulation of yeast 4E-BP, Caf20, by TORC1.
- To determine if TORC1 influences Caf20 phosphorylation and ribosome association.
- To elucidate the role of the TORC1-4E-BP pathway in yeast translational control.
Main Methods:
- In vivo analysis of Caf20 phosphorylation status.
- In vitro kinase assays to assess direct TORC1 phosphorylation of Caf20.
- Biochemical assays to detect TORC1-dependent Caf20 recruitment to ribosomes.
Main Results:
- TORC1 influences the phosphorylation of Caf20 in vivo.
- TORC1 directly phosphorylates Caf20 in vitro.
- TORC1 mediates the recruitment of Caf20 to the 80S ribosome.
Conclusions:
- Yeast 4E-BP, Caf20, is a direct substrate of TORC1.
- TORC1-mediated phosphorylation and ribosome recruitment of Caf20 suggest its role in translation initiation.
- This study clarifies the involvement of the TORC1-4E-BP axis in regulating yeast translational efficiency.
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