Figure 1.
UBC9 is a new partner of Aurora-A. (A) UBC9 and Aurora-A interact in vitro. Recombinant GST–UBC9 protein previously incubated with or without Aurora-A(His)6 was incubated with nickel–agarose beads (Ni–NTA–agarose), whereas Aurora-A (His)6 recombinant protein was incubated in the presence of GST–UBC9 or GST alone and loaded onto glutathione-sepharose. The presence of Aurora-A and UBC9 was tested by blotting using anti-Aurora-A and anti-UBC9 antibodies. (B) Aurora-A and UBC9 interact in vivo. Anti-AurkA antibodies were able to immunoprecipitate UBC9 (left panels) and anti-UBC9 antibodies immunoprecipitated Aurora-A proteins (middle panels) from HeLa cell protein extracts. The same reactions performed with beads only (left lines) or control IgG (right panels) were unable to immunoprecipitate either UBC9 or Aurora-A.