Cooperative Gsx2-DNA Binding Requires DNA Bending and a Novel Gsx2 Homeodomain Interface.

Autor:	Webb JA; Department of Molecular and Cellular Biosciences, University of Cincinnati College of Medicine, Cincinnati, OH 45267, USA., Farrow E; Graduate Program in Molecular and Developmental Biology, Cincinnati Children's Hospital Research Foundation, Cincinnati, OH 45229, USA.; Medical-Scientist Training Program, University of Cincinnati College of Medicine, Cincinnati, OH 45229, USA., Cain B; Division of Developmental Biology, Cincinnati Children's Hospital Medical Center, 3333 Burnet Ave, MLC 7007, Cincinnati, OH 45229, USA., Yuan Z; Department of Molecular and Cellular Biosciences, University of Cincinnati College of Medicine, Cincinnati, OH 45267, USA., Yarawsky AE; Division of Immunobiology, Cincinnati Children's Hospital Medical Center, 3333, Burnet Ave, Cincinnati, OH 45229, USA., Schoch E; Department of Medical Education, University of Cincinnati College of Medicine, Cincinnati, OH 45229, USA., Gagliani EK; Department of Chemistry, Xavier University, Cincinnati, OH 45207, USA., Herr AB; Division of Immunobiology, Cincinnati Children's Hospital Medical Center, 3333, Burnet Ave, Cincinnati, OH 45229, USA., Gebelein B; Division of Developmental Biology, Cincinnati Children's Hospital Medical Center, 3333 Burnet Ave, MLC 7007, Cincinnati, OH 45229, USA.; Department of Pediatrics, University of Cincinnati College of Medicine, Cincinnati, OH 45229, USA., Kovall RA; Department of Molecular and Cellular Biosciences, University of Cincinnati College of Medicine, Cincinnati, OH 45267, USA.
Jazyk:	angličtina
Zdroj:	BioRxiv : the preprint server for biology [bioRxiv] 2023 Dec 08. Date of Electronic Publication: 2023 Dec 08.
DOI:	10.1101/2023.12.08.570805
Abstrakt:	The conserved Gsx homeodomain (HD) transcription factors specify neural cell fates in animals from flies to mammals. Like many HD proteins, Gsx factors bind A/T-rich DNA sequences prompting the question - how do HD factors that bind similar DNA sequences in vitro regulate specific target genes in vivo ? Prior studies revealed that Gsx factors bind DNA both as a monomer on individual A/T-rich sites and as a cooperative homodimer to two sites spaced precisely seven base pairs apart. However, the mechanistic basis for Gsx DNA binding and cooperativity are poorly understood. Here, we used biochemical, biophysical, structural, and modeling approaches to (1) show that Gsx factors are monomers in solution and require DNA for cooperative complex formation; (2) define the affinity and thermodynamic binding parameters of Gsx2/DNA interactions; (3) solve a high-resolution monomer/DNA structure that reveals Gsx2 induces a 20° bend in DNA; (4) identify a Gsx2 protein-protein interface required for cooperative DNA binding; and (5) determine that flexible spacer DNA sequences enhance Gsx2 cooperativity on dimer sites. Altogether, our results provide a mechanistic basis for understanding the protein and DNA structural determinants that underlie cooperative DNA binding by Gsx factors, thereby providing a deeper understanding of HD specificity. Competing Interests: Declaration of Interests R.K. is on the scientific advisory board of Cellestia Biotech AG and has received research funding from Cellestia for projects unrelated to this manuscript. A.B.H. serves on the scientific advisory board for Hoth Therapeutics, Inc., and holds equity in Hoth Therapeutics and Chelexa BioSciences, LLC. The remaining authors declare no competing interests.
Databáze:	MEDLINE
Externí odkaz:	Zobrazit plný text záznamu