10.1371/journal.ppat.1007515.g008 John V. Dzimianski John V. Dzimianski Brianna S. Beldon Brianna S. Beldon Courtney M. Daczkowski Courtney M. Daczkowski Octavia Y. Goodwin Octavia Y. Goodwin Florine E. M. Scholte Florine E. M. Scholte Éric Bergeron Éric Bergeron Scott D. Pegan Scott D. Pegan Second site of FARV vOTU interaction with di-Ub. Public Library of Science 2019 novel X-ray crystal structures Crimean-Congo hemorrhagic fever virus Recent genomic characterization ISG 15 fluorogenic substrates Hughes orthonairovirus species Nairoviridae family vOTU substrate preferences DUB deubiquitinase activity Post-translational modification gene product 15 nairovirus genomic diversity tumor domain protease 2019-01-10 19:09:59 Figure https://plos.figshare.com/articles/figure/Second_site_of_FARV_vOTU_interaction_with_di-Ub_/7575236 <p>(A) Model of FARV vOTU (reddish orange) bound to K63-linked di-Ub (purple; PDB ID 2JF5). FARV vOTU was overlaid with CCHFV vOTU bound to Ub (PDB ID 3PRP; not rendered) based on secondary structure alignment of the vOTUs. The distal Ub was anchored to the bound mono-Ub by aligning the secondary structure in Coot, followed by manual bond rotations within Lys63 of the proximal Ub in PyMol to model a plausible fit with minimal clashes based on the CCHFV vOTU active site and protease surface. The predicted region of FARV vOTU engagement with the proximal Ub is indicated by a black box. (B) Closeup view of the predicted region, with the residues selected for mutation shown as sticks. Activity of the mutants relative to WT is shown for Ub-AMC, Ub-Rh110, K48 di-Ub FRET-TAMRA, and K63 di-Ub FRET-TAMRA (right).</p>