SIX3 and SIX6 interact with GEMININ via C-terminal regions
The histoarchitecture and function of eye and forebrain depend on a well-controlled balance between cell proliferation and differentiation. For example, the binding of the cell cycle regulator GEMININ to CDT1, which is a part of the pre-replication complex, promotes cell differentiation. Homeodomain...
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| Format: | Article |
| Language: | English |
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Elsevier
2019-12-01
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| Series: | Biochemistry and Biophysics Reports |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2405580819301736 |
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| author | Diana C. Turcu Johan R. Lillehaug Hee-Chan Seo |
| author_facet | Diana C. Turcu Johan R. Lillehaug Hee-Chan Seo |
| author_sort | Diana C. Turcu |
| collection | DOAJ |
| description | The histoarchitecture and function of eye and forebrain depend on a well-controlled balance between cell proliferation and differentiation. For example, the binding of the cell cycle regulator GEMININ to CDT1, which is a part of the pre-replication complex, promotes cell differentiation. Homeodomain transcription factors SIX3 and SIX6 also interact with GEMININ of which SIX3-GEMININ interaction promotes cell proliferation, whereas the nature of SIX6-GEMININ interaction has not been studied to date. We investigated SIX3/SIX6 and GEMININ interactions using bimolecular fluorescence complementation, surface plasmon resonance and isothermal titration calorimetry. Interactions between SIX3/SIX6 and GEMININ were detected in mammalian cells in culture. The presence of the C-terminal regions of SIX3 and SIX6 proteins, but not their SIX domains or homeodomains as previously thought, were required for interaction with GEMININ. Interestingly, the disordered C- and N- terminal regions of GEMININ were involved in binding to SIX3/SIX6. The coiled-coil region of GEMININ, which is the known protein-binding domain and also interacts with CDT1, was not involved in GEMININ-SIX3/SIX6 interaction. Using SPR and ITC, SIX3 bound GEMININ with a micromolar affinity and the binding stoichiometry was 1:2 (SIX3 - GEMININ). The present study gives new insights into the binding properties of SIX proteins, especially the role of their variable and disordered C-terminal regions. Keywords: SIX3, SIX6, GEMININ, Cell-cycle regulation, Bimolecular fluorescence complementation (BiFC), Surface plasmon resonance (SPR), Isothermal titration calorimetry (ITC), Protein interaction |
| first_indexed | 2024-12-21T11:54:37Z |
| format | Article |
| id | doaj.art-d082616e83b04d879b9460b4d187c241 |
| institution | Directory Open Access Journal |
| issn | 2405-5808 |
| language | English |
| last_indexed | 2024-12-21T11:54:37Z |
| publishDate | 2019-12-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Biochemistry and Biophysics Reports |
| spelling | doaj.art-d082616e83b04d879b9460b4d187c2412022-12-21T19:04:59ZengElsevierBiochemistry and Biophysics Reports2405-58082019-12-0120SIX3 and SIX6 interact with GEMININ via C-terminal regionsDiana C. Turcu0Johan R. Lillehaug1Hee-Chan Seo2Department of Biological Sciences, University of Bergen, Bergen, NorwayDepartment of Biological Sciences, University of Bergen, Bergen, NorwayCorresponding author.; Department of Biological Sciences, University of Bergen, Bergen, NorwayThe histoarchitecture and function of eye and forebrain depend on a well-controlled balance between cell proliferation and differentiation. For example, the binding of the cell cycle regulator GEMININ to CDT1, which is a part of the pre-replication complex, promotes cell differentiation. Homeodomain transcription factors SIX3 and SIX6 also interact with GEMININ of which SIX3-GEMININ interaction promotes cell proliferation, whereas the nature of SIX6-GEMININ interaction has not been studied to date. We investigated SIX3/SIX6 and GEMININ interactions using bimolecular fluorescence complementation, surface plasmon resonance and isothermal titration calorimetry. Interactions between SIX3/SIX6 and GEMININ were detected in mammalian cells in culture. The presence of the C-terminal regions of SIX3 and SIX6 proteins, but not their SIX domains or homeodomains as previously thought, were required for interaction with GEMININ. Interestingly, the disordered C- and N- terminal regions of GEMININ were involved in binding to SIX3/SIX6. The coiled-coil region of GEMININ, which is the known protein-binding domain and also interacts with CDT1, was not involved in GEMININ-SIX3/SIX6 interaction. Using SPR and ITC, SIX3 bound GEMININ with a micromolar affinity and the binding stoichiometry was 1:2 (SIX3 - GEMININ). The present study gives new insights into the binding properties of SIX proteins, especially the role of their variable and disordered C-terminal regions. Keywords: SIX3, SIX6, GEMININ, Cell-cycle regulation, Bimolecular fluorescence complementation (BiFC), Surface plasmon resonance (SPR), Isothermal titration calorimetry (ITC), Protein interactionhttp://www.sciencedirect.com/science/article/pii/S2405580819301736 |
| spellingShingle | Diana C. Turcu Johan R. Lillehaug Hee-Chan Seo SIX3 and SIX6 interact with GEMININ via C-terminal regions Biochemistry and Biophysics Reports |
| title | SIX3 and SIX6 interact with GEMININ via C-terminal regions |
| title_full | SIX3 and SIX6 interact with GEMININ via C-terminal regions |
| title_fullStr | SIX3 and SIX6 interact with GEMININ via C-terminal regions |
| title_full_unstemmed | SIX3 and SIX6 interact with GEMININ via C-terminal regions |
| title_short | SIX3 and SIX6 interact with GEMININ via C-terminal regions |
| title_sort | six3 and six6 interact with geminin via c terminal regions |
| url | http://www.sciencedirect.com/science/article/pii/S2405580819301736 |
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