Fig. 1. Myosin VI interacts with the replisome.

a Schematic representation of myosin VI (adapted from Magistrati and Polo⁴⁰) showing the positions of the ubiquitin-binding MIU and MyUb domains (blue) adjacent to the cargo-binding domain (CBD, green). The amino acid sequence shows a triple-Lys repeat involved in DNA binding¹⁸ (orange box) and the WWY motif (green box), a well-characterized protein interaction site. The three-helix bundle at the N-terminal tail is indicated in red. Amino acid numbering is according to the short isoform (isoform 2). b Set-up of the SILAC experiment for identification of MyUb interaction partners. c GO term analysis (GO biological process) of proteins identified to interact with the MyUb domain (fold change > 4, FDR < 0.05) using EnrichR. d Volcano plot of protein groups identified in the SILAC interactome experiment. Mean log₂ fold change of all replicates between GST-MyUb and GST are plotted against the −log₁₀ FDR. Significantly enriched proteins are shown in red (fold change > 2, FDR < 0.05). Interactors involved in DNA replication and repair are highlighted and labeled. e Validation of selected candidates by pulldown assays from total cell lysates with recombinant GST-MyUb, followed by western blotting and Ponceau S staining. Results were confirmed by at least two independent experiments. Source data are provided as a Source Data file.