AMPK and AKT protein kinases hierarchically phosphorylate the N-terminus of the FOXO1 transcription factor, modulating interactions with 14-3-3 proteins

Maria Saline, Lukas Badertscher, Madita Wolter, Roxanne Lau, Anders Gunnarsson, Tomas Jacso, Tyrrell Norris, Christian Ottmann, Arjan Snijder (Corresponding author)

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Abstract

Forkhead box protein O1 (FOXO1) is a transcription factor involved in various cellular processes such as glucose metabolism, development, stress resistance, and tumor suppression. FOXO1's transcriptional activity is controlled by different environmental cues through a myriad of posttranslational modifications. In response to growth factors, the serine/threonine kinase AKT phosphorylates Thr24 and Ser256 in FOXO1 to stimulate binding of 14-3-3 proteins, causingFOXO1inactivation. In contrast, low nutrient and energy levels induce FOXO1 activity. AMP-activated protein kinase (AMPK), a master regulator of cellular energy homeostasis, partly mediates this effect through phosphorylation of Ser383 and Thr649 in FOXO1. In this study, we identified Ser22 as an additional AMPK phosphorylation site in FOXO1's N terminus, with Ser22 phosphorylation preventing binding of 14-3-3 proteins. The crystal structure of a FOXO1 peptide in complex with 14-3-3 σ at 2.3 Å resolution revealed that this is a consequence of both steric hindrance and electrostatic repulsion. Furthermore, we found that AMPK-mediated Ser22 phosphorylation impairs Thr24 phosphorylation by AKT in a hierarchical manner. Thus, numerous mechanisms maintain FOXO1 activity via AMPK signaling. AMPK-mediated Ser22 phosphorylation directly and indirectly averts binding of 14-3-3 proteins, whereas phosphorylation of Ser383 and Thr649 complementarily stimulates FOXO1 activity. Our results shed light on a mechanism that integrates inputs from both AMPK and AKT signaling pathways in a small motif to fine-tune FOXO1 transcriptional activity.

Original languageEnglish
Pages (from-to)13106-13116
Number of pages11
JournalJournal of Biological Chemistry
Volume294
Issue number35
DOIs
Publication statusPublished - 30 Aug 2019

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14-3-3 Proteins
Phosphorylation
AMP-Activated Protein Kinases
Protein Kinases
Transcription Factors
Forkhead Transcription Factors
Protein-Serine-Threonine Kinases
Post Translational Protein Processing
Static Electricity
Metabolism
Electron energy levels
Nutrients
Cues
Tumors
Electrostatics
Intercellular Signaling Peptides and Proteins
Homeostasis
Crystal structure
Glucose
Food

Cite this

Saline, Maria ; Badertscher, Lukas ; Wolter, Madita ; Lau, Roxanne ; Gunnarsson, Anders ; Jacso, Tomas ; Norris, Tyrrell ; Ottmann, Christian ; Snijder, Arjan. / AMPK and AKT protein kinases hierarchically phosphorylate the N-terminus of the FOXO1 transcription factor, modulating interactions with 14-3-3 proteins. In: Journal of Biological Chemistry. 2019 ; Vol. 294, No. 35. pp. 13106-13116.
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title = "AMPK and AKT protein kinases hierarchically phosphorylate the N-terminus of the FOXO1 transcription factor, modulating interactions with 14-3-3 proteins",
abstract = "Forkhead box protein O1 (FOXO1) is a transcription factor involved in various cellular processes such as glucose metabolism, development, stress resistance, and tumor suppression. FOXO1's transcriptional activity is controlled by different environmental cues through a myriad of posttranslational modifications. In response to growth factors, the serine/threonine kinase AKT phosphorylates Thr24 and Ser256 in FOXO1 to stimulate binding of 14-3-3 proteins, causingFOXO1inactivation. In contrast, low nutrient and energy levels induce FOXO1 activity. AMP-activated protein kinase (AMPK), a master regulator of cellular energy homeostasis, partly mediates this effect through phosphorylation of Ser383 and Thr649 in FOXO1. In this study, we identified Ser22 as an additional AMPK phosphorylation site in FOXO1's N terminus, with Ser22 phosphorylation preventing binding of 14-3-3 proteins. The crystal structure of a FOXO1 peptide in complex with 14-3-3 σ at 2.3 {\AA} resolution revealed that this is a consequence of both steric hindrance and electrostatic repulsion. Furthermore, we found that AMPK-mediated Ser22 phosphorylation impairs Thr24 phosphorylation by AKT in a hierarchical manner. Thus, numerous mechanisms maintain FOXO1 activity via AMPK signaling. AMPK-mediated Ser22 phosphorylation directly and indirectly averts binding of 14-3-3 proteins, whereas phosphorylation of Ser383 and Thr649 complementarily stimulates FOXO1 activity. Our results shed light on a mechanism that integrates inputs from both AMPK and AKT signaling pathways in a small motif to fine-tune FOXO1 transcriptional activity.",
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AMPK and AKT protein kinases hierarchically phosphorylate the N-terminus of the FOXO1 transcription factor, modulating interactions with 14-3-3 proteins. / Saline, Maria; Badertscher, Lukas; Wolter, Madita; Lau, Roxanne; Gunnarsson, Anders; Jacso, Tomas; Norris, Tyrrell; Ottmann, Christian; Snijder, Arjan (Corresponding author).

In: Journal of Biological Chemistry, Vol. 294, No. 35, 30.08.2019, p. 13106-13116.

Research output: Contribution to journalArticleAcademicpeer-review

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T1 - AMPK and AKT protein kinases hierarchically phosphorylate the N-terminus of the FOXO1 transcription factor, modulating interactions with 14-3-3 proteins

AU - Saline, Maria

AU - Badertscher, Lukas

AU - Wolter, Madita

AU - Lau, Roxanne

AU - Gunnarsson, Anders

AU - Jacso, Tomas

AU - Norris, Tyrrell

AU - Ottmann, Christian

AU - Snijder, Arjan

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