Extended Data Fig. 7: Conservation of C6 in orthopoxviruses.
From: TRIM5α restricts poxviruses and is antagonized by CypA and the viral protein C6
a) Amino acid sequence alignment of orthopoxvirus C6 proteins created by Clustal Omega. Amino acids were colour-coded according to their properties. Asterisks (*) indicate fully conserved residues, colons (:) indicate amino acids with strongly similar properties and dots (.) indicate amino acids with weakly similar properties. Numbers on the left indicate % amino acid identity compared to VACV WR C6. b) Orthopoxvirus C6 co-precipitation with endogenous TRIM5α in T-REx-293 cells inducibly expressing TAP-tagged C6 proteins from VACV WR, rabbitpox virus (RPXV), cowpox virus strain Brighton Red (CPXV), elephantpox virus, camelpox virus (CMLV) strain CMS, monkeypox virus (MPXV) strain UK2022, MPXV Zaire and variola major virus (VARV) strain India 1967. TAP-tagged proteins were affinity-purified using Strep-Tactin beads. c) Immunoblot showing TRIM5α down-regulation in HEK293T cells during RPXV and elephantpox virus infections. Cells were infected at 5 PFU/cell for the indicated times. MG132 was added at 2 hpi. In b), cells were treated with 150 ng/mL doxycycline for 16 h before harvest. Inputs and AP proteins in b) or protein extracts in c) were analysed by SDS-PAGE and immunoblotted for epitope/protein. α-Actin, and C6 and D8 were controls for loading and viral infection, respectively. Data shown are representative of two independent experiments.
