FIGURE 1.

RhoGDIα is lysine-acetylated. A, left, overview of reported lysine acetylation sites of RhoGDIα identified in recent quantitative proteomic screens in cells and tissues as indicated. Right, localization of the eight acetylated lysine residues in the RhoA·RhoGDIα structure (PDB entry 4F38). Lys-99, Lys-105, Lys-127, Lys-138, Lys-141, and Lys-178 are in the immunoglobulin domain, and Lys-43 and Lys-52 are in the N-terminal domain. Yellow, RhoA; gray, RhoGDIα. Acetylated lysines are highlighted in red. B, RhoGDIα is endogenously acetylated in HEK293T cells, as shown by immunoprecipitation of lysine acetylated proteins and probing of RhoGDIα. Acetylation is regulated by KDACs, as seen by the increase of acetylated RhoGDIα after incubating the cells with KDAC inhibitors for 6 h. IB for α-tubulin serves as control. +, with KDAC inhibitors; −, without KDAC inhibitors. C, transiently expressed His₆-tagged RhoGDIα is acetylated in HEK293T cells, as shown by Ni²⁺-NTA pull-down (PD) and IB. The Ac-Lys signal increased upon KDAC inhibitor treatment (+). Anti-His IB serves as loading control.