Extended Data Fig. 3: VV A51R associates with microtubules (MTs) and tethers hSpt16SUMO to MTs.
From: FEAR antiviral response pathway is independent of interferons and countered by poxvirus proteins
a, Alignment of VV A51R with Proline Rich (PR) Region and Region 1 of human Tau40 MT-binding domains (MTBD)25. “KXGS” motif conserved in all four MTBDs in Tau is boxed. Residues in red indicate those converted to alanine in the triple A51R mutant. b, immunofluorescence staining of U2OS cells transfected with wild-type or triple mutant FA51R expression constructs after 24 h. Where indicated, 40 μM nocodazole was added to depolymerize MTs 6 h post-transfection MTs. Scale bar, 5 μm. c, Overview of in vitro MT co-sedimentation assay. d-e, Stain-free protein gels showing in vitro MT co-sedimentation assays with purified wild-type and mutant His-A51R. S, supernatant; P, pellet. f, Co-IP of transfected wild-type and mutant FA51R constructs with hSpt16 and tubulin in 293 T WCE. g, Overview of in vitro Flag-hSpt16 Co-IP with tubulin in the absence/presence of His-A51R. h, in vitro Co-IP of purified Flag-hSpt16, His-A51R and tubulin. Images in b, d-f, and h are representative of at least two independent experiments. Images in c and g were created with biorender.com.
