Extended Data Fig. 4: Identification of TRIM5α and CypA interaction partner(s).
From: TRIM5α restricts poxviruses and is antagonized by CypA and the viral protein C6
a) L3 co-precipitation with CypA WT and mutants. T-REx-293 TRIM5-/-CypA-/- cells were transfected with TAP-tagged TRIM5α, CypA WT +/− CsA or CypA mutants, R55A and F113A, and infected with vΔC6 at 3 PFU/cell for 12 h. b) L3 dimerisation and direct interaction with CypA and TRIM5α shown using cell-free transcription and translation system. TAP-tagged L3, TRIM5α WT, ΔSPRY and CypA were co-expressed with HA-tagged L3. In a) and b) TAP-tagged proteins were precipitated using Strep-Tactin. Inputs and AP proteins were analysed by SDS-PAGE and immunoblotted for the indicated epitope/protein. α-Actin and D8 were controls for loading and viral infection, respectively. Data from a) are representative of three independent experiments, and b) is from two.
