Figure 4.

LRRK2 interacts with and phosphorylates Futsch. A–D, hLRRK2 physically interacts with Futsch. A, Extracts from Da–GAL4>hLRRK2–Flag or control animals were immunoprecipitated with mAb 22C10 against Futsch, and the presence of hLRRK2–Flag in the immunocomplex was detected by Western blot with anti-hLRRK2. B, IP of extracts as described in A was performed to pull down hLRRK2–Flag with anti-Flag. The presence of Futsch was detected by Western blot with mAb 22C10. C, Genetic interaction between dLRRK and futsch. D, Futsch coexpression rescues synaptic defects caused by presynaptic overexpression of hLRRK2. E, hLRRK2 phosphorylates Futsch in vitro. In vitro kinase assays were performed with substrate only or substrate plus WT, G2019S, or KD forms of hLRRK2. Top, Autoradiography; bottom, Western blots as control for protein loading. Blots were probed with anti-Futsch and anti-hLRRK2.