Contrasting DNA-binding behaviour by ISL1 and LHX3 underpins differential gene targeting in neuronal cell specification
| Autor: | Ann H. Kwan, Ngaio C. Smith, Jill Trewhella, Jacqueline M. Matthews, Lorna Wilkinson-White |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
QH301-705.5
Cell LIM-homeodomain transcription factors Article 03 medical and health sciences chemistry.chemical_compound Homeodomain-DNA interaction Structural Biology Gene expression medicine Surface plasmon resonance Biology (General) Transcription factor 030304 developmental biology ComputingMethodologies_COMPUTERGRAPHICS DNA-binding specificity 0303 health sciences Chemistry 030302 biochemistry & molecular biology Gene targeting SAXS Cell biology Transcriptional complex medicine.anatomical_structure ISL1 LHX3 DNA |
| Zdroj: | Journal of Structural Biology: X, Vol 5, Iss, Pp 100043-(2021) Journal of Structural Biology: X |
| ISSN: | 2590-1524 |
| Popis: | Graphical abstract Highlights • The mechanisms by which ISL1 and LHX3 specify neuronal cell identity are unknown. • EMSA/SPR data show ISL1 and LHX3 have markedly different DNA-binding behaviours. • SAXS shows ISL1/LHX3:DNA complexes are flexible in nature. • ISL1 binds DNA poorly but appears to modulate the DNA-binding specificity of LHX3. The roles of ISL1 and LHX3 in the development of spinal motor neurons have been well established. Whereas LHX3 triggers differentiation into interneurons, the additional expression of ISL1 in developing neuronal cells is sufficient to redirect their developmental trajectory towards spinal motor neurons. However, the underlying mechanism of this action by these transcription factors is less well understood. Here, we used electrophoretic mobility shift assays (EMSAs) and surface plasmon resonance (SPR) to probe the different DNA-binding behaviours of these two proteins, both alone and in complexes mimicking those found in developing neurons, and found that ISL1 shows markedly different binding properties to LHX3. We used small angle X-ray scattering (SAXS) to structurally characterise DNA-bound species containing ISL1 and LHX3. Taken together, these results have allowed us to develop a model of how these two DNA-binding modules coordinate to regulate gene expression and direct development of spinal motor neurons. |
| Databáze: | OpenAIRE |
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