Anthrax Edema Factor Toxicity Is Strongly Mediated by the N-end Rule.docVIP

Anthrax Edema Factor Toxicity Is Strongly Mediated by the N-end Rule Clinton E. Leysath, Damilola D. Phillips, Devorah Crown, Rasem J. Fattah, Mahtab Moayeri, Stephen H. Leppla * Laboratory of Parasitic Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland, United States of America Abstract Anthrax edema factor (EF) is a calmodulin-dependent adenylate cyclase that converts adenosine triphosphate (ATP) into 3’–5’-cyclic adenosine monophosphate (cAMP), contributing to the establishment of Bacillus anthracis infections and the resulting pathophysiology. We show that EF adenylate cyclase toxin activity is strongly mediated by the N- end rule, and thus is dependent on the identity of the N-terminal amino acid. EF variants having different N-terminal residues varied by more than 100-fold in potency in cultured cells and mice. EF variants having unfavorable, destabilizing N-terminal residues showed much greater activity in cells when the E1 ubiquitin ligase was inactivated or when proteasome inhibitors were present. Taken together, these results show that EF is uniquely affected by ubiquitination and/or proteasomal degradation. Citation: Leysath CE, Phillips DD, Crown D, Fattah RJ, Moayeri M, et al. (2013) Anthrax Edema Factor Toxicity Is Strongly Mediated by the N-end Rule. PLoS ONE 8(8): e74474. doi:10.1371/journal.pone.0074474 Editor: Prosper N Boyaka, The Ohio State University, United States of America Received June 19, 2013; Accepted July 30, 2013; Published August 28, 2013 This is an open-access article, free of all copyright, and may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. The work is made available under the Creative Commons CC0 public domain dedication. Funding: This research was supported by the Intramural Research Program of the National Institute of Allergy and Infect