Fig 1. Nmo phosphorylates Pk.

(A) Prickle is phosphorylated by Nmo kinase but not dROK or Hpo kinases. Band shift assay of in vitro translated Pk (common region), Pan/dTCF and Myosin binding subunit (Mbs). The open arrowhead denotes the size of the unphosphorylated form (compared to no kinase lane). Note the band shift of Pk. Pan and Mbs serve as positive and negative controls, respectively. (B) Schematic of Pk constructs used in this study. The Pk protein scheme is shown above with the Pk-Sple N terminus (blue), short Pk N terminus as black line, PET domain (green) and three LIM domains (yellow). Constructs as denoted here are indicated by thick black lines: common (sequence shared between Pk and Pk-Sple isoforms); C-terminus; N-Δ1; N-Δ2 (both C-terminal deletions as indicated); C1; C2; Dom (PET and LIM domains); Sple N-terminus (Pk-Sple specific N-terminal extension); PET domain; M (Middle sequence) (Methods for sequence details). Full lines indicate fragments that are phosphorylated by Nmo, dashed lines indicate unphosphorylated fragments. (C) In vitro kinase assay gel using purified Nmo kinase and in vitro translated Pk fragments (from panel B). Upper panel; radiograph showing phosphorylation; autophosphorytion of Nmo is denoted by *N. Vang C-term is used as negative control (also [24]). Common, C-term, N-Δ1, N-Δ2, and M fragments are phosphorylated by Nmo. Corresponding Coomassie-stained gel is shown below—full-length fragments of individual constructs are indicated by *.