Fig. 2: US31 gene expression

a NCBI Blast homology analysis of the US31 gene using the Toledo strain as a reference sequence. Blue represents identity with the reference strain whereas red represents differences. PnHV_2: Panine herpesvirus 2; HHV_5: Human herpesvirus 5; Rh_CMV: Rhesus Cytomegalovirus; MaLe_CMV: Mandrillus leucophaeus cytomegalovirus; CeHV_5: Cercopithecine herpesvirus 5; PaUrCMV: Papio ursinus cytomegalovirus; CeHV: Cercopithecine herpesvirus 5; CyMaCMV_ot/ma: Cynomolgus macaque cytomegalovirus strain Ottawa/Mauritius. b mRNA detection of the US31 gene in HEK293T and COS-7 cells after infection/ transfection with Ad-US31 or Ad-GFP (NC, control) or pcDNA3.1/US31 vector (Figures do not show) for 24 h. Expression was normalized to that of GAPDH. c Western blot analysis of US31 protein expression in HEK293T and COS-7 cells 24 h after infection/ transfection with Ad-US31 or Ad-GFP (NC) or pcDNA3.1/US31 (Figures do not show). d Analysis of US31 expression in Ad-US31- or Ad-GFP (control)-infected THP-1-derived macrophages (48 h). Red staining indicates US31 expression (performed using a rabbit antibody against US31); blue staining indicates nuclei (DAPI). e Output of Expasy functional analysis of US31. f Sequence alignment of the UBR domain in Ubr1 E3 ligases from different organisms (Sc, Saccharomyces cerevisiae; Hs, Homo sapiens; Mm, Mus musculus; Kl, Kluyveromyces lactis; Sp, Schizosaccharomyces pombe). Shading indicates residues that are identical (red) or highly conserved (yellow) in all sequences. Filled circles indicate residues that coordinate the zinc ions (orange for Zn1, green for Zn2). Key determinants for binding of the N-terminal residue of the substrate are marked with blue triangles and critical residues identified by previous genetic screening are indicated with purple triangles below the sequence³⁹. g Predicted 3-dimensional protein structure of US31. The red area represents the nuclear localization signal sequence