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Published on: November 9, 2017
A Phosphomimetic Study Implicates Ser557 in Regulation of FOXP2 DNA Binding
Ashleigh Blane1, Heini W Dirr1, Sylvia Fanucchi2
1Protein Structure-Function Research Unit, School of Molecular and Cell Biology, University of the Witwatersrand, Johannesburg, 2050, South Africa.
Phosphorylation of Ser557 in the FOXP2 forkhead domain (FHD) disrupts DNA binding. This suggests a regulatory role for phosphorylation in FOXP2-mediated transcription control.
Area of Science:
- Molecular Biology
- Genetics
- Biochemistry
Background:
- FOXP2 is a crucial transcription factor during embryonic development.
- It binds DNA via its forkhead domain (FHD) recognition helix.
- Serine 557 (Ser557) within this helix is a potential phosphorylation site.
Purpose of the Study:
- To investigate the impact of Ser557 phosphorylation on FOXP2 FHD structure and DNA binding.
- To compare the behavior of wild-type FOXP2 FHD with a phosphomimetic mutant (S557E).
Main Methods:
- Construction and analysis of the S557E phosphomimetic mutant.
- Electrophoretic mobility shift assays (EMSAs) to assess DNA binding.
- Molecular docking simulations to compare protein-DNA interactions.
Main Results:
- The S557E mutation did not alter secondary or tertiary protein structure.
- Phosphorylation significantly reduced the propensity of FOXP2 FHD to form dimers.
- DNA binding affinity was markedly decreased in the S557E mutant compared to wild-type.
- Molecular docking revealed fewer favorable DNA interactions for the phosphomimetic and phosphorylated forms.
Conclusions:
- Phosphorylation of Ser557 likely disrupts FOXP2 FHD DNA binding through electrostatic and steric effects.
- This phosphorylation event may serve as a regulatory mechanism controlling FOXP2 transcriptional activity.
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