A novel fic (filamentation induced by cAMP) protein from clostridium difficile reveals an inhibitory motif-independent adenylylation/AMPylation mechanism

Emil Dedic, Husam Alsarraf, Ditte Hededam Welner, Ole Østergaard, Oleg I. Klychnikov, Paul J. Hensbergen, Jeroen Corver, Hans C. Van Leeuwen, René Jørgensen*

*Corresponding author

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review


Filamentation induced by cAMP (Fic) domain proteins have been shown to catalyze the transfer of the AMP moiety from ATP onto a protein target. This type of post-translational modification was recently shown to play a crucial role in pathogenicity mediated by two bacterial virulence factors. Herein we characterize a novel Fic domain protein that we identified from the human pathogen Clostridium difficile. The crystal structure shows that the protein adopts a classical all-helical Fic fold, which belongs to class II of Fic domain proteins characterized by an intrinsic N-terminal autoinhibitory ô-helix. A conserved glutamate residue in the inhibitory helix motif was previously shown in other Fic domain proteins to prevent proper binding of the ATP ô-phosphate. However, here we demonstrate that both ATP binding and autoadenylylation activity of the C. difficile Fic domain protein are independent of the inhibitory motif. In support of this, the crystal structure of a mutant of this Fic protein in complex with ATP reveals that the ô-phosphate adopts a conformation unique among Fic domains that seems to override the effect of the inhibitory helix. These results provide important structural insight into the adenylylation reaction mechanism catalyzed by Fic domains. Our findings reveal the presence of a class II Fic domain protein in the human pathogen C. difficile that is not regulated by autoinhibition and challenge the current dogma that all class I-III Fic domain proteins are inhibited by the inhibitory ô-helix.

TidsskriftJournal of Biological Chemistry
Udgave nummer25
Sider (fra-til)13286-13300
Antal sider15
StatusUdgivet - 17 jun. 2016
Udgivet eksterntJa

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© 2016 by The American Society for Biochemistry and Molecular Biology, Inc.

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