FIG. 2.

Oligomerization of onco-Dbl is homophilic in nature and is mediated by the DH domain. (A) Complex formation of GST-Dbl with the HA-tagged Dbl family members and DH-PH chimeras. HA-tagged Dbl, Ost, Lbc, TrioN, TrioC, and the HA-tagged DH domain-PH domain chimeras made between Dbl and Ost (Dbl/Ost), between Dbl and Lbc (Dbl/Lbc), and between Lbc and Dbl (Lbc/Dbl) were expressed in Cos-7 cells. The cell lysates were incubated with glutathione-agarose-immobilized GST-Dbl for 30 min. The GST-Dbl coprecipitates, as well as the input cell lysates, were subjected to anti-HA Western blotting analysis. (B) The DH domain is responsible for complex formation with GST-Dbl. Cell lysates expressing the HA-DH domain of Dbl were subjected to a GST-Dbl, GST-PH, or GST pull-down assay. The input lysates and the glutathione-agarose coprecipitates were visualized by an anti-HA Western blot.