Functional dissection of Timekeeper ( Tik ) implicates opposite roles for CK2 and PP2A during Drosophila neurogenesis

Repression by E(spl)M8 during inhibitory Notch (N) signaling (lateral inhibition) is regulated, in part, by protein kinase CK2, but the involvement of a phosphatase has been unclear. The studies we report here employ Tik , a unique dominant‐negative (DN) mutation in the catalytic subunit of CK2, in a Gal4‐ UAS based assay for impaired lateral inhibition. Specifically, overexpression of Tik elicits ectopic bristles in N + flies and suppresses the retinal defects of the gain‐of‐function allele N spl . Functional dissection of the two substitutions in Tik (M 161 K and E 165 D), suggests that both mutations contribute to its DN effects. While the former replacement compromises CK2 activity by impairing ATP‐binding, the latter affects a conserved motif implicated in binding the phosphatase PP2A. Accordingly, overexpression of microtubule star ( mts ), the PP2A catalytic subunit closely mimics the phenotypic effects of loss of CK2 functions in N + or N spl flies, and elicits notched wings, a characteristic of N mutations. Our findings suggest antagonistic roles for CK2 and PP2A during inhibitory N signaling. genesis 47:647–658, 2009. © 2009 Wiley‐Liss, Inc.