Journal: bioRxiv
Article Title: DDIAS is a single-stranded DNA-binding effector of the TOPBP1-CIP2A complex in mitosis
doi: 10.1101/2025.09.09.675193
Figure Lengend Snippet: (A) Schematic showing the layout of conserved domains and motifs in TOPBP1. Numbered boxes represent BRCT domains, with phospho-peptide binding domains in green and domains lacking phospho-peptide binding activity in grey. Names of known TOPBP1 binding partners are shown below the domains they interact with. CBR, CIP2A-binding region; AAD, ATR-activation domain. (B) Sequence alignments of the two TOPBP1 BRCT7-binding motifs in DDIAS and a known interaction partner, FANCJ. (C) Sequence alignments of the TOPBP1 BRCT5-binding motif in DDIAS and a known interaction partner, BLM. (D) AlphaFold 3 predictions showing conserved phospho-peptide motifs in DDIAS in complex with TOPBP1 BRCT domains.
Article Snippet: The following antibodies were used at the indicated dilutions: BLM (A300-110A, Bethyl Laboratories, 1/2000), BRCA1 (sc-6954, Santa Cruz Biotechnology, 1/200), BRCA2 (OP95, Merck, 1/2000), CIP2A (14805, Cell Signaling Technology, 1/1000), FANCJ (4578, Cell Signaling Technology, 1/500), GFP (11814460001, Roche, 1/5000), H2AX (NB100-383, Novus Biologicals, 1/5000), HaloTag (G9211, Promega, 1/1000), PLK1 (05-844, Merck, 1/4000), RAD9 (sc-8324, Santa Cruz Biotechnology, 1/1000), RPA2 (ab10359, Abcam, 1/10,000), TOPBP1 (A300-111A, Bethyl Laboratories, 1/1000).
Techniques: Binding Assay, Activity Assay, Activation Assay, Sequencing