Journal article
Direct imaging of the recruitment and phosphorylation of S6K1 in the mTORC1 pathway in living cells
- Abstract:
- Knowledge of protein signalling pathways in the working cell is seen as a primary route to identifying and developing targeted medicines. In recent years there has been a growing awareness of the importance of the mTOR pathway, making it an attractive target for therapeutic intervention in several diseases. Within this pathway we have focused on S6 kinase 1 (S6K1), the downstream phosphorylation substrate of mTORC1, and specifically identify its juxtaposition with mTORC1. When S6K1 is co-expressed with raptor we show that S6K1 is translocated from the nucleus to the cytoplasm. By developing a novel biosensor we demonstrate in real-time, that phosphorylation and de-phosphorylation of S6K1 occurs mainly in the cytoplasm of living cells. Furthermore, we show that the scaffold protein raptor, that typically recruits mTOR substrates, is not always involved in S6K1 phosphorylation. Overall, we demonstrate how FRET-FLIM imaging technology can be used to show localisation of S6K1 phosphorylation in living cells and hence a key site of action of inhibitors targeting mTOR phosphorylation.
- Publication status:
- Published
- Peer review status:
- Peer reviewed
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(Preview, Version of record, pdf, 2.4MB, Terms of use)
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- Publisher copy:
- 10.1038/s41598-019-39410-z
Authors
- Publisher:
- Nature Research
- Journal:
- Scientific Reports More from this journal
- Volume:
- 9
- Issue:
- 2019
- Article number:
- 3408
- Publication date:
- 2019-03-04
- Acceptance date:
- 2019-01-23
- DOI:
- ISSN:
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2045-2322
- Pmid:
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30833605
- Language:
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English
- Keywords:
- Pubs id:
-
pubs:980873
- UUID:
-
uuid:14443750-f647-40a4-8f83-65b0bfd3a599
- Local pid:
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pubs:980873
- Source identifiers:
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980873
- Deposit date:
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2019-04-03
- ARK identifier:
Terms of use
- Copyright holder:
- Ahmed et al
- Copyright date:
- 2019
- Notes:
- © The Author(s) 2019. This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
- Licence:
- CC Attribution (CC BY)
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