FIGURE 1:

Rga6 is a Rho2 and Cdc42 GAP. (A) Schematic representation of Rga6 protein, indicating the serine-rich region (SR), GAP domain, and polybasic region (PBR) at the C-terminus. (B) β-Galactosidase analysis of the two-hybrid interaction between rga6⁺ and the constitutively active versions of rho1, rho2, rho3, rho4, and cdc42 lacking the coding sequence of the three C-terminal amino acids. (C, D) Levels of total and GTP-bound Cdc42 (C) and Rho2 (D) in rga6Δ strain. Extracts from wild-type and rga6Δ cells expressing HA-cdc42⁺, and HA-rho2⁺ were pulled down with GST-CRIB or GST-C21RBD, respectively, and blotted with anti-HA antibody (middle). Total HA-Rho proteins in the extracts were visualized by Western blot using anti-HA antibody (top). Actin was visualized in the same extracts as loading control (bottom). The ratio of GTP-bound to total protein is represented as a bar graph and shows the average and SD of four different experiments for each GTPase.