Assay and properties of rab6 interaction with dynein-dynactin complexes.

RAB GTPases help to maintain the fidelity of membrane trafficking events by recruiting cytosolic tethering and motility factors to vesicle and organelle membranes. In the case of Rab6, it recruits the dynein-dynaction complex to Golgi-associated vesicles via an adaptor protein of the Bicaudal-D family. Here we describe methods for the identification of Rab6-binding partners in cell extracts. We then focus on the biochemical analysis of interactions with the dynein-dynactin complex and the adaptor proteins Bicaudal-D1 and -D2. Standard protocols for yeast two-hybrid analysis, and biochemical assays for the analysis of the interactions between Rab6, Bicaudal-D, and the subunits of the dynein-dynactin complex are outlined.