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The forkhead domain hinge-loop plays a pivotal role in DNA binding and transcriptional activity of FOXP2

  • Gavin Morris , Stoyan Stoychev , Previn Naicker , Heini W. Dirr and Sylvia Fanucchi EMAIL logo
Published/Copyright: June 7, 2018
Biological Chemistry
From the journal Biological Chemistry Volume 399 Issue 8

Abstract

Forkhead box (FOX) proteins are a ubiquitously expressed family of transcription factors that regulate the development and differentiation of a wide range of tissues in animals. The FOXP subfamily members are the only known FOX proteins capable of forming domain-swapped forkhead domain (FHD) dimers. This is proposed to be due to an evolutionary mutation (P539A) that lies in the FHD hinge loop, a key region thought to fine-tune DNA sequence specificity in the FOX transcription factors. Considering the importance of the hinge loop in both the dimerisation mechanism of the FOXP FHD and its role in tuning DNA binding, a detailed investigation into the implications of mutations within this region could provide important insight into the evolution of the FOX family. Isothermal titration calorimetry and hydrogen exchange mass spectroscopy were used to study the thermodynamic binding signature and changes in backbone dynamics of FOXP2 FHD DNA binding. Dual luciferase reporter assays were performed to study the effect that the hinge-loop mutation has on FOXP2 transcriptional activity in vivo. We demonstrate that the change in dynamics of the hinge-loop region of FOXP2 alters the energetics and mechanism of DNA binding highlighting the critical role of hinge loop mutations in regulating DNA binding characteristics of the FOX proteins.

Keywords: backbone dynamics; DNA-binding; electrostatics; forkhead; FOXP; hinge-loop

Funding source: National Research Foundation

Award Identifier / Grant number: 80681 to S.F., 68898 to H.W.D.

Funding statement: This work was supported by the University of the Witwatersrand; South African National Research Foundation, Funder Id: 10.13039/501100001321 (Grant 80681 to S.F., 68898 to H.W.D.); the South African Research Chairs Initiative of the Department of Science and Technology (Grant 64788 to H.W.D.) and the Medical Research Council of South Africa, Funder Id: 10.13039/501100001342.

Acknowledgements

We would like to acknowledge Kerry Hanmer and Dr. Demetra Mavri-Damelin from the School of Molecular and Cell Biology at the University of the Witwatersrand for their assistance in culturing the HEK293 cells used in the dual luciferase reporter assays. We also extend our gratitude to Dr. Sonja Vernes (Max Planck Institute for Psycholinguistics, The Netherlands) for the generous gift of the pcDNA4-FOXP2 vector used in the dual luciferase reporter assays.

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Supplementary Material:

The online version of this article offers supplementary material (https://doi.org/10.1515/hsz-2018-0185).

Received: 2018-03-15
Accepted: 2018-04-26
Published Online: 2018-06-07
Published in Print: 2018-07-26

©2018 Walter de Gruyter GmbH, Berlin/Boston

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https://doi.org/10.1515/hsz-2018-0185
Keywords for this article
backbone dynamics; DNA-binding; electrostatics; forkhead; FOXP; hinge-loop

Articles in the same Issue

  1. Frontmatter
  2. Reviews
  3. Exploratory cell dynamics: a sense of touch for cells?
  4. Kallikrein-related peptidases and associated microRNAs as promising prognostic biomarkers in gastrointestinal malignancies
  5. Sphingolipid metabolism – an ambiguous regulator of autophagy in the brain
  6. Minireview
  7. Insulin-like signaling within and beyond metazoans
  8. Research Articles/Short Communications
  9. Genes and Nucleic Acids
  10. Oxidation of 1-N2-etheno-2′-deoxyguanosine by singlet molecular oxygen results in 2′-deoxyguanosine: a pathway to remove exocyclic DNA damage?
  11. Protein Structure and Function
  12. Novel approach to quorum quenching: rational design of antibacterials in combination with hexahistidine-tagged organophosphorus hydrolase
  13. The forkhead domain hinge-loop plays a pivotal role in DNA binding and transcriptional activity of FOXP2
  14. New clues into the self-assembly of Vmh2, a basidiomycota class I hydrophobin
  15. Membranes, Lipids, Glycobiology
  16. Model construction of Niemann-Pick type C disease in zebrafish
  17. Cell Biology and Signaling
  18. Upregulation of Twist is involved in Gli1 induced migration and invasion of hepatocarcinoma cells
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