Figure 1. CK2 interacts with the LOV-1 and PC-1 PLAT domain and enriches in the cilia of male specific neurons. (a) Anatomy of C. elegans male and PKD-2-expressing neurons. Top, DIC side view image of an adult C. elegans male oriented head (left) to ventral up tail (right). Bottom,
lov-1 and
pkd-2 are expressed in male head CEM neurons (left) and tail ray RnB and hook HOB neurons (right). The cilium, dendrite,cell body, and axon of the CEMD (dorsal CEM) and CEMV (ventral CEM) neuron are drawn in the male head diagram. CEMD and CEMV are arranged as left-right pairs (one side shown here). Cilia are in the nose region (dashed rectangular box). The pharynx is green. Positions of nuclei of all
lov-1 and
pkd-2 expressing cells in the C. elegans adult male tail, modified from (Sulston et al., 1980). Ray neurons are arranged as left-right bilateral pairs (ventral up view shown here). The male tail has nine bilaterally arranged rays (numbered 1-9, anterior to posterior)required for response and turning behaviors (Liu and Sternberg, 1995). HOB is an asymmetric ciliated hook neuron that mediates vulva location behavior (Liu and Sternberg, 1995). For simplicity, only the dendrite of R3B left and HOB is shown. The R3B cilium is indicated by a dashed rectangular box. (b) A GST pull down assay demonstrates a direct in vitro interaction between KIN-10 and the LOV-1 PLAT domain.In vitro-translated 35S-labeled KIN-10 protein was incubated with GST or GST-PLAT (amino acids 2112-2302). Autoradiography shows35S-labeled KIN-10 protein is specifically retained in the GST-PLAT column. A Western blot developed with an anti-GST antibody detects GST and GST-PLAT. (c) Y2H mapping reveals the CK2 signature domain of KIN-10 mediates interaction with the PLAT domains of humanPC-1 and C. elegans LOV-1. Different
kin-10 fragments were cloned into the Y2H AD vector pGADT7. The C. elegans and human PLAT domain(amino acids 3164-3349) were engineered into the Y2H BD vector pGBKT7. Y2H interactions were accessed by growth ability on SD-Leu-Trp-His-Ade plates. (d) Confocal micrographs of Ppkd-2::PKD-2::GFP, Ppkd-2::KIN-3::GFP (using
pkd-2 promoter to drive KIN-3::GFP fusion protein expression) and Ppkd-2::KIN-10::GFP (using
pkd-2 promoter to drive KIN-10::GFP fusion protein expression) transgenic males. PKD-2 localizes to cilia and cell bodies (excluding nuclei) of male-specific neurons. KIN-3 and KIN-10 are localized and enriched in the cilia of the male-specific RnB, HOB and CEM neurons. KIN-3 and KIN-10 also localize weakly in dendrites and axons and strongly in cell bodies (especially in nuclei). Dashed rectangular boxes show the ciliary zone of the CEM neurons. Dashed circles show the ciliary zone of HOB neuron. Dashed diamond boxes show the ciliary zone of RnB neuron in ray 3. Labeled bars indicate length in micrometer (um).