a p-loop mutation in gα subunits prevents transition to the active state implications for g-protein signaling in fungal pathogenesis在gαp-loop突变子单元防止过渡到真菌致病蛋白信号的活动状态的影响.pdfVIP

A P-loop Mutation in Ga Subunits Prevents Transition to the Active State: Implications for G-protein Signaling in Fungal Pathogenesis 1. 1.¤ 2,3 1 Dustin E. Bosch , Francis S. Willard *, Ravikrishna Ramanujam , Adam J. Kimple , Melinda D. Willard1¤, Naweed I. Naqvi2,3,4, David P. Siderovski1,5* 1 Department of Pharmacology, The University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, United States of America, 2 Fungal Patho-Biology Group, Temasek Life Sciences Laboratory, Singapore, 3 School of Biological Sciences, Nanyang Technological University, Singapore, 4 Department of Biological Sciences, National University of Singapore, Singapore, 5 UNC Neuroscience Center and Lineberger Comprehensive Cancer Center, The University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, United States of America Abstract Heterotrimeric G-proteins are molecular switches integral to a panoply of different physiological responses that many organisms make to environmental cues. The switch from inactive to active Gabc heterotrimer relies on nucleotide cycling by the Ga subunit: exchange of GTP for GDP activates Ga, whereas its intrinsic enzymatic activity catalyzes GTP hydrolysis to GDP and inorganic phosphate, thereby reverting Ga to its inactive state. In several genetic studies of filamentous fungi, such as the rice blast fungus Magnaporthe oryzae, a G42R mutation in the phosphate-binding loop of Ga subunits is assumed to be GTPase-deficient and thus constitutively active. Here, we demonstrate that Ga(G42R) mutants are not GTPase deficient, but rather incapable of achieving the activated conformation. Two crystal structure models suggest that Arg-42 prevents a typical switch region conformational change