The Fic protein Doc uses an inverted substrate to phosphorylate and inactivate EF-Tu.

TitleThe Fic protein Doc uses an inverted substrate to phosphorylate and inactivate EF-Tu.
Publication TypeJournal Article
Year of Publication2013
AuthorsCastro-Roa, D., Garcia-Pino A., De Gieter S., van Nuland N. A. J., Loris R., and Zenkin N.
JournalNat Chem Biol
Volume9
Issue12
Pagination811-7
Date Published2013 Dec
Type of Articleta
ISSN1552-4469
KeywordsEscherichia coli Proteins, Gene Expression Regulation, Bacterial, Guanosine Triphosphate, Models, Molecular, Mutation, Nucleotidyltransferases, Peptide Elongation Factor Tu, Phosphorylation, Protein Folding, RNA, Transfer
Abstract

Fic proteins are ubiquitous in all of the domains of life and have critical roles in multiple cellular processes through AMPylation of (transfer of AMP to) target proteins. Doc from the doc-phd toxin-antitoxin module is a member of the Fic family and inhibits bacterial translation by an unknown mechanism. Here we show that, in contrast to having AMPylating activity, Doc is a new type of kinase that inhibits bacterial translation by phosphorylating the conserved threonine (Thr382) of the translation elongation factor EF-Tu, rendering EF-Tu unable to bind aminoacylated tRNAs. We provide evidence that EF-Tu phosphorylation diverged from AMPylation by antiparallel binding of the NTP relative to the catalytic residues of the conserved Fic catalytic core of Doc. The results bring insights into the mechanism and role of phosphorylation of EF-Tu in bacterial physiology as well as represent an example of the catalytic plasticity of enzymes and a mechanism for the evolution of new enzymatic activities.

DOI10.1038/nchembio.1364
Alternate JournalNat. Chem. Biol.
PubMed ID24141193
PubMed Central IDPMC3836179
Grant List202994 / / European Research Council / International
BB/J006378/1 / / Biotechnology and Biological Sciences Research Council / United Kingdom
/ / Biotechnology and Biological Sciences Research Council / United Kingdom
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