Wee1 localization to nodes requires Cdr2 kinase activity and the Wee1 N terminus. (A) Schematic of Wee1 and Cdr2 functional domains. Values represent amino acid positions. (B) The Wee1 N terminus interacts with WT Cdr2 but not with kinase-dead Cdr2(E177A) in the yeast two-hybrid assay. Transformants were selected on a double-dropout (DDO) plate, and interactions were tested on a quadruple-dropout plate containing aureobasidin, X-gal, and 3-AT (QDO/A/X/3AT) plate. Positive interactions are indicated by growth of blue colonies on selective plates. (C) Localization of the indicated Wee1 constructs overexpressed from the P81nmt1 promoter. Middle–focal plane widefield images with inverted contrast are shown, and insets show enlarged views of dashed boxes. Bars, 5 µm. (D) Length of dividing septated cells of the indicated genotypes (means ± SD; n > 50 cells). ****, P < 0.0001; n.s., P > 0.05.