direct activation of rhoa by reactive oxygen species requires a redox-sensitive motif直接激活rhoa活性氧需要redox-sensitive图案.pdfVIP

Direct Activation of RhoA by Reactive Oxygen Species Requires a Redox-Sensitive Motif 1. 1. 2 1,3 Amir Aghajanian *, Erika S. Wittchen , Sharon L. Campbell , Keith Burridge 1 Department of Cell and Developmental Biology, University of North Carolina at Chapel Hill, Lineberger Comprehensive Cancer Center, Chapel Hill, North Carolina, United States of America, 2 Department of Biochemistry and Biophysics, University of North Carolina at Chapel Hill, Lineberger Comprehensive Cancer Center, Chapel Hill, North Carolina, United States of America, 3 UNC McAllister Heart Institute, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, United States of America Abstract Background: Rho family GTPases are critical regulators of the cytoskeleton and affect cell migration, cell-cell adhesion, and cell-matrix adhesion. As with all GTPases, their activity is determined by their guanine nucleotide-bound state. Understanding how Rho proteins are activated and inactivated has largely focused on regulatory proteins such as guanine nucleotide exchange factors (GEFs) and GTPase activating proteins (GAPs). However, recent in vitro studies have indicated that GTPases may also be directly regulated by redox agents. We hypothesized that this redox-based mechanism occurs in cells and affects cytoskeletal dynamics, and in this report we conclude this is indeed a novel mechanism of regulating the GTPase RhoA. Methodology/Principal Findings: In this report, we show that RhoA can be directly activated by reactive oxygen species (ROS) in cells, and that this requires two critical cysteine residues located in a unique redox-sensitive motif within the phosphoryl binding loop. First, we show that ROS can reve