Fig. 4.

Identification and characterization of TMEM205 as oHSV-interacting host cell protein. A Protein complexes directly interacting with oHSV viral particles were isolated by IP using anti-oHSV gC Ab, separated by SDS-PAGE and visualized by Coomassie blue staining. Arrowheads point to some of the proteins differentially identified by mass spectrometry. B The WB of the IP samples was probed for human TMEM205. Anti-oHSV gC Ab from oHSV was used for IP. The interaction was further confirmed for OVTOKO and JHOC cells with IP: TMEM205 and WB: gC (lower panel). UT is untreated control; T1 is C-terminal labelled TMEM205 antibody and T2 is N-terminal labelled antibody for both the sets. C Wild type OVTOKO cells, TMEM 205 knockdown, and OV TMEM scrambled control cell lines (TM Si) were infected with oHSV to study viral binding and entry. Cells were processed and viral DNA was quantified by qPCR. Data represent the mean ± SEM of three independent experiments. D Ubiquitination assay showing ample degradation of TMEM205 mediated by ubiquitin post treatment with oHSV in the first 2 lanes. Thus, oHSV helps in degradation of TMEM205 via the proteasome pathway