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. 2020 May 11;9:e53789. doi: 10.7554/eLife.53789

Figure 3. Conservation of the pUL7:pUL51 interaction across herpesviruses.

(A–D) HEK 293 T cells were co-transfected with GFP-tagged pUL7 homologues from human herpesviruses, or with GFP alone, and with mCherry tagged pUL51 homologues. Cells were lysed 24 hr post-transfection and incubated with anti-GFP (A, B, D) or anti-RFP (C) resin to capture protein complexes before being subjected to SDS-PAGE and immunoblotting using the antibodies shown. All immunoblots are representative of at least two independent experiments performed by different scientists. (A) mCherry-tagged homologues of pUL51 are captured by GFP-pUL7 homologues, but not by GFP alone. (B) GFP-pUL7 co-precipitates with pUL51 (HSV-1) and pORF7 (VZV), but not with pUL71 (HCMV) or pORF55 (KSHV). (C) pUL51-mCherry co-precipitates with pUL7 but not with homologues from other herpesviruses. (D) The VZV pUL7 homologue pORF53 co-precipitates with VZV pORF7, but not with pUL51 homologues from other herpesviruses. (E) Molecular surfaces of the pUL7 and pUL51 core heterodimer, colored by residue conservation across the α-herpesviruses. Residues that mediate the pUL7:pUL51 interaction are outlined.

Figure 3.

Figure 3—figure supplement 1. pUL51 does not co-precipitate pUL14 in uninfected cultured cells.

Figure 3—figure supplement 1.

(A) Core heterodimer of pUL7:pUL51 with residues required for the reported pUL51 and pUL14 (Oda et al., 2016) highlighted in pink. Inset shows pUL7 and the first helix of pUL51 as surfaces, and side chains of residues that were substituted with alanine in Oda et al., 2016 are shown as sticks. (B) HEK 293 T cells were co-transfected with myc-tagged pUL14 from HSV-1 along with GFP-pUL7 and pUL51-mCherry, as shown. Co-immunoprecipitation of myc-pUL14 with pUL51-mCherry is not observed either in the presence or absence of pUL7. Image shown is representative of three independent experiments.