Morris, GStoychev, Stoyan HNaicker, PrevinDirr, HWFanucchi, Stephanie2019-03-222019-03-222018-07Morris, G. et al. 2018. The forkhead domain hinge-loop plays a pivotal role in DNA binding and transcriptional activity of FOXP2. Biological Chemistry, vol. 399(8): 881-8931431-67301437-4315https://www.degruyter.com/view/j/bchm.ahead-of-print/hsz-2018-0185/hsz-2018-0185.xmlDOI: https://doi.org/10.1515/hsz-2018-0185http://hdl.handle.net/10204/10813Copyright: 2018 De GruyterForkhead 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.enDNA-bindingFOXPBackbone dynamicsElectrostaticsForkheadHinge-loopArticleMorris, G., Stoychev, S. H., Naicker, P., Dirr, H., & Fanucchi, S. (2018). The forkhead domain hinge-loop plays a pivotal role in DNA binding and transcriptional activity of FOXP2. http://hdl.handle.net/10204/10813Morris, G, Stoyan H Stoychev, Previn Naicker, HW Dirr, and Stephanie Fanucchi "The forkhead domain hinge-loop plays a pivotal role in DNA binding and transcriptional activity of FOXP2." (2018) http://hdl.handle.net/10204/10813Morris G, Stoychev SH, Naicker P, Dirr H, Fanucchi S. The forkhead domain hinge-loop plays a pivotal role in DNA binding and transcriptional activity of FOXP2. 2018; http://hdl.handle.net/10204/10813.TY - Article AU - Morris, G AU - Stoychev, Stoyan H AU - Naicker, Previn AU - Dirr, HW AU - Fanucchi, Stephanie AB - 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. DA - 2018-07 DB - ResearchSpace DP - CSIR KW - DNA-binding KW - FOXP KW - Backbone dynamics KW - Electrostatics KW - Forkhead KW - Hinge-loop LK - https://researchspace.csir.co.za PY - 2018 SM - 1431-6730 SM - 1437-4315 T1 - The forkhead domain hinge-loop plays a pivotal role in DNA binding and transcriptional activity of FOXP2 TI - The forkhead domain hinge-loop plays a pivotal role in DNA binding and transcriptional activity of FOXP2 UR - http://hdl.handle.net/10204/10813 ER -