Ubiquitination and Degradation of the Hominoid-Specific Oncoprotein TBC1D3 Is Mediated by CUL7 E3 Ligase
Figure 2
(A) In vitro transcription/translation assay. Plasmid inserts encoding putative TBC1D3 binding partners were transcribed and translated in vitro using a coupled, T7 polymerase-driven reaction with 35S-methionine. The SARA and Rab5a are the full-length human proteins while the CUL7 is an amino-terminal portion of the human CUL7. The proteins were resolved using SDS-PAGE, and TBC1D3 binding was visualized by autoradiography. The migration of molecular weight standards is indicated. (B) TBC1D3 co-immunoprecipitates with CUL7. HeLa cells were transfected with plasmids encoding HA-CUL7 and Myc-TBC1D3. Cells were lysed and immunoprecipitated with anti-Myc antibody. The immunoprecipitates were resolved by SDS-PAGE and blotted with anti-HA and anti-Myc antibodies.
