Transfer of HTLV-1 p8 and Gag to target T-cells depends on VASP, a novel interaction partner of p8
Fig 2
The EVH1-domain and the F- and G-actin binding domains of VASP are required for the interaction with p8.
(A) Domain structure of VASP protein and mutants. (B) 293T cells were transfected with expression plasmids p8-HA and FLAG-VASP, FLAG-VASPΔEVH1, FLAG-VASP-EVH1 or the respective empty control vectors. After 48 h, cells were lysed and 10% of the lysates were taken as input (IN). Co-immunoprecipitations (IPs) were performed using anti-FLAG antibodies. Immunoblots are shown. HC, heavy chain. One representative out of four independent experiments is shown. (C) Densitometry was performed to quantitate the amount of p8-HA after precipitation and values were normalized on the respective FLAG-VASP expression in the input. p8 binding to VASP wildtype was set to 100%. Bars indicate the means of four independent experiments ± SE and values were compared to VASP wildtype using Student’s t-test. * indicates p<0.05. (D) 293T cells were transfected with expression plasmids p8-HA and FLAG-VASP, FLAG-VASPΔPP, FLAG-VASPΔGAB, FLAG-VASPΔFAB, FLAG-VASPΔD, FLAG-VASPΔC or FLAG-EVH2. IPs were performed as described in (A). One representative out of four independent experiments is shown.