Fig. 3: Vpr forms a ternary complex with VprBP WD40-AR and Plk4 CPB. | Nature Communications

Fig. 3: Vpr forms a ternary complex with VprBP WD40-AR and Plk4 CPB.

From: Centrosome amplification and aneuploidy driven by the HIV-1-induced Vpr•VprBP•Plk4 complex in CD4+ T cells

Fig. 3: Vpr forms a ternary complex with VprBP WD40-AR and Plk4 CPB.

Coimmunoprecipitation analyses performed with HEK293T cells transfected with the indicated constructs. Asterisk in (a), degradation product; Black triangles in (c), Increased amounts of Vpr DNA transfected; arrows in (d), mGFP-STREP-containing ligands; numbers in (b–d), relative signal intensities. e SEC profile and SDS-PAGE analysis demonstrating the purification of the DDB1•Vpr•VprBP WD40-AR(1057–1507)•Plk4 CPB(581–808) complex. Arrows, respective proteins loaded in each lane; red numbers, an estimated binding stoichiometry approximated from the Coomassie Brilliant Blue–stained protein intensity. Note that the level of coprecipitating VprBP is proportional to that of Vpr expressed in the lysates. f Interferometric scattering mass spectrometry (iSCAMS) data showing the Vpr•VprBP WD40-AR•Plk4 CPB complex forming within 1 min after mixing all components at 200 mM NaCl. The complex is sensitive to 400 mM NaCl (the 4th panel). The 11-kDa Vpr, which binds tightly to VprBP, cannot be detected due to its small particle size. Red arrow, the Vpr•VprBP WD40-AR complex (red asterisk, a presumed dimer); yellow arrow, Plk4 CPB; thick black arrow, the Vpr•VprBP WD40-AR•Plk4 CPB complex.

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