9XS1

Crystal structure of FOXM1 DNA binding domain to specific dsDNA substrate

Summary for 9XS1

• Entry DOI: 10.2210/pdb9xs1/pdb
• Descriptor: Forkhead box protein M1, DNA (5'-D(*TP*TP*TP*GP*TP*TP*TP*AP*TP*TP*TP*GP*TP*TP*TP*GP*TP*TP*TP*AP*TP*TP*TP*G)-3'), DNA (5'-D(*CP*AP*AP*AP*TP*AP*AP*AP*CP*AP*AP*AP*CP*AP*AP*AP*TP*AP*AP*AP*CP*AP*AP*A)-3'), ... (6 entities in total)
• Functional Keywords: transcription factor, dna-binding domain, dna binding protein/dna, dna binding protein-dna complex
• Biological source: Homo sapiens (human); More
• Total number of polymer chains: 7
• Total formula weight: 78527.90

Authors

Lv, M.Q.,Sun, M.X.,Zhou, W.W. (deposition date: 2025-11-20, release date: 2026-04-15)

Primary citation

Sun, M.,Wang, L.,Cui, J.,Zhang, L.,Shi, Y.,Xu, C.,Zhou, W.,Lv, M.
Structural basis for the FOXM1 DNA binding domain to specific dsDNA substrate.
Acta Biochim.Biophys.Sin., 2026

PubMed Abstract: Forkhead box protein M1 (FOXM1) is a key transcription factor that regulates cell cycle progression and is frequently overexpressed in human cancers, driving tumor proliferation and therapy resistance. FOXM1 recognizes the canonical forkhead response element (FKH motif, RYAAAYA) through its conserved DNA-binding domain (DBD). Here, we report the high-resolution crystal structure of the FOXM1-DBD in complex with a double-stranded DNA substrate containing two FKH motifs. The structure reveals that FOXM1-DBD adopts the canonical winged-helix fold, with the third α-helix (α3) inserted into the DNA major groove to mediate sequence-specific recognition. Within this helix, Asn283, Arg286, and His287 form an essential triad that engages DNA bases through specific hydrogen bonds and hydrophobic interactions. Using structure-guided mutagenesis of key DNA-interacting residues combined with biophysical validation by isothermal titration calorimetry (ITC) and DNA binding assessment via electrophoretic mobility shift assay (EMSA), we confirm the functional importance of these residues and uncover position-dependent tolerance to base substitutions within the FKH motif. Furthermore, we demonstrate that FOXM1 overexpression promotes cell proliferation and upregulates the transcription of target genes in a DBD-dependent manner. Our findings provide a structural basis for understanding the DNA recognition mechanism of FOXM1 and offer mechanistic insights into how FOXM1 selectively binds to its genomic targets to regulate transcription.

PubMed: 41889283
DOI: 10.3724/abbs.2026036

Experimental method

X-RAY DIFFRACTION (2.5 Å)