Figure 9.

MTV1 and AGD5 Interact with CHC.

(A) Domain structure of MTV1 with the N-terminal ENTH domain, a putative clathrin binding box LIDTG, and 6 DxF (DLL) motifs implicated in clathrin binding.

(B) and (C) Pull-down experiments. GST-tagged bait proteins were bound to sepharose beads and incubated with Col-0 total protein extract. Flow-through was collected (FT), beads were washed extensively, and bound material (eluate) was analyzed by immunoblot using anti-CHC antibody. Asterisks in loading control mark the bait proteins. Clathrin binds GST-MTV1 full-length protein (B); clathrin binding occurs via the C-terminal part of the protein (C).

(D) NEV/AGD5 interacts with CHC in Tap-tag experiments. Left, gel separation of copurified proteins with AGD5-TAP from Arabidopsis cells in two independent experiments; asterisks mark bait protein and arrowheads the position of CHC. Right, table of mass spectrometry data from both experiments for peptides corresponding to AGD5 and CHC (see Supplemental Table 2 online).

[See online article for color version of this figure.]