Fig. 1.

Doxycycline-inducible expression of the VP1-2 USP domain. (a) Summary of various N-terminal constructs of VP1-2, NT1, NT2, NT3, NT5, and NT6 in relation to general features of VP1-2 organization. Numbering refers to the final residue in each construct. The N-terminal core USP is indicated by brown shading, the NLS in pink shading, the central core of VP1-2 in gray, and the C-terminal highly conserved region in blue. Various alignments within each of the alpha, beta, and gamma classes indicate this overall organization, as summarized in Fig. S1 in the supplemental material. Previously characterized regions involved in UL37, VP16, and UL25 binding are indicated (7, 14, 24, 26, 30, 31, 40). (b) Induction of NT1 and NT1.C65A after transfection in 293-Tet On cells. Cells were transfected with the appropriate vectors, Dox was added 24 h later, and cells were harvested 48 h after Dox addition. The primary NT1 species are indicated by an arrow. The panel in the right hand side shows a longer exposure revealing additional lower-abundance species and the presence of NT1* in NT1.C65A, a species never observed for NT1.