Fig. 2.

The rod^Z3 mutation changes a conserved glycine in the Rod_C domain, but does not affect RZZ assembly. (A) The mutation in rod^Z3 results in a G to E change at residue 1973 in a conserved C-terminal region of the protein. A Clustal alignment around the mutated region of Rod from mammals and dipterans shows that G1973 (red box) has been conserved since the divergence of mosquitoes and flies. The corresponding position in mammalian Rod is always a polar residue. (B) Levels of Rod, Zw10 and Zwilch are similar in extracts of WT and rod^Z3-derived syncytial embryos, and immunoprecipitation (IP) of Rod brings down Zw10 and Zwilch equally well in both WT and rod^Z3 extracts. Immunoblots are for Rod, Zw10 and Zwilch in input lysates of WT or rod^Z3 derived embryos (left) (Tubulin is a loading control). Immunoprecipitates using anti-Rod, (middle), or control non-immune rabbit serum are shown on the right). The lower molecular mass in the Zwilch blots is an unrelated contaminant. (C) RZZ from WT and rod^Z3-derived syncytial embryos both form high-molecular-mass complexes. Fractions of a sucrose density gradient centrifugation of embryo extracts immunoblotted for Rod. Arrows indicate migration of size markers.