Fig. 7: Identifying sequence determinants for host-virus interactions in IDRs.

a Workflow for selecting putative short linear interaction sites in IDRs from AF-Multimer models. b Comparison of the residue length for the putative interaction motif to the confidence of the predicted interface (average pLDDT of all amino acids within the selected site). c Host proteins cross-linked to UL12, depicted on the monomeric AF2 model of UL12, color coded by pLDDT. d Predicted MAPK8-UL12 dimer with a putative D-motif highlighted magenta. The inset shows the magnified region of the D-motif in UL12 interacting with the D-site in MAPK8. Shown below the inset is a solved crystal structure of MAPK8 with the D-motif containing peptide of NFAT4 (pdb 2xrw). e Mutational disruption of the D-motif in UL12 in the viral genome of HSV-1, creating P39A, L44A-UL12. Label-free comparative AP-MS of the wildtype UL12 and P39A, L44A-U12 interactomes. The p-values are based on two-sided t tests without multiple hypothesis correction of n = 3 replicates. Boxed regions highlight proteins that interact significantly stronger with P39A, L44A-UL12 (shaded green) or WT-UL12 (shaded orange), based on a cut-off at P = 0.01 and log2 fold-change of 4. f Mutational disruption of the 14-3-3 binding site in UL12 in the viral genome of HSV-1, creating S41A, P43A-UL12. SILAC-based comparative AP-MS of the wildtype UL12 to the S41A, P43A mutated UL12 interactome, with both (n = 2, label-swap) replicates depicted. Boxed regions highlight proteins that interact stronger with S41A, P43A-UL12 (shaded green) or WT-UL12 (shaded orange), based on a log2 fold-change cut-off of one in both replicates. Source data are provided.