title: Intrinsic regulation of FIC-domain AMP-transferases by oligomerization and automodification.
abstract: Filamentation induced by cyclic AMP (FIC)-domain enzymes catalyze adenylylation or other posttranslational modifications of target proteins to control their function. Recently, we have shown that Fic enzymes are autoinhibited by an α-helix (αinh) that partly obstructs the active site. For the single-domain class III Fic proteins, the αinh is located at the C terminus and its deletion relieves autoinhibition. However, it has remained unclear how activation occurs naturally. Here, we show by structural, biophysical, and enzymatic analyses combined with in vivo data that the class III Fic protein NmFic from Neisseria meningitidis gets autoadenylylated in cis, thereby autonomously relieving autoinhibition and thus allowing subsequent adenylylation of its target, the DNA gyrase subunit GyrB. Furthermore, we show that NmFic activation is antagonized by tetramerization. The combination of autoadenylylation and tetramerization results in nonmonotonic concentration dependence of NmFic activity and a pronounced lag phase in the progress of target adenylylation. Bioinformatic analyses indicate that this elaborate dual-control mechanism is conserved throughout class III Fic proteins.
authors.affiliation: Focal Area Structural Biology and Biophysics, Biozentrum, University of Basel, CH-4056 Basel, Switzerland; Focal Area Infection Biology, Biozentrum, University of Basel, CH-4056 Basel, Switzerland;
authors.forename: Frédéric V
authors.initials: FV
authors.lastname: Stanger
authors.affiliation: Focal Area Structural Biology and Biophysics, Biozentrum, University of Basel, CH-4056 Basel, Switzerland;
authors.forename: Björn M
authors.identifier: http://orcid.org/0000-0002-3135-7964
authors.initials: BM
authors.lastname: Burmann
authors.affiliation: Focal Area Infection Biology, Biozentrum, University of Basel, CH-4056 Basel, Switzerland;
authors.forename: Alexander
authors.initials: A
authors.lastname: Harms
authors.affiliation: Focal Area Structural Biology and Biophysics, Biozentrum, University of Basel, CH-4056 Basel, Switzerland;
authors.forename: Hugo
authors.initials: H
authors.lastname: Aragão
authors.affiliation: Research IT, Biozentrum, University of Basel, CH-4056 Basel, Switzerland;
authors.forename: Adam
authors.initials: A
authors.lastname: Mazur
authors.affiliation: Biophysics Facility, Biozentrum, University of Basel, CH-4056 Basel, Switzerland.
authors.forename: Timothy
authors.initials: T
authors.lastname: Sharpe
authors.affiliation: Focal Area Infection Biology, Biozentrum, University of Basel, CH-4056 Basel, Switzerland; christoph.dehio@unibas.ch sebastian.hiller@unibas.ch tilman.schirmer@unibas.ch.
authors.forename: Christoph
authors.initials: C
authors.lastname: Dehio
authors.affiliation: Focal Area Structural Biology and Biophysics, Biozentrum, University of Basel, CH-4056 Basel, Switzerland; christoph.dehio@unibas.ch sebastian.hiller@unibas.ch tilman.schirmer@unibas.ch.
authors.forename: Sebastian
authors.identifier: http://orcid.org/0000-0002-6709-4684
authors.initials: S
authors.lastname: Hiller
authors.affiliation: Focal Area Structural Biology and Biophysics, Biozentrum, University of Basel, CH-4056 Basel, Switzerland; christoph.dehio@unibas.ch sebastian.hiller@unibas.ch tilman.schirmer@unibas.ch.
authors.forename: Tilman
authors.initials: T
authors.lastname: Schirmer
mesh_terms: D001426:Bacterial Proteins / Q000378:metabolism*; D001704:Biopolymers / Q000378:metabolism*; D000242:Cyclic AMP / Q000378:metabolism*; D027081:DNA Gyrase / Q000378:metabolism; D008958:Models, Molecular; D009345:Neisseria meningitidis / Q000201:enzymology*; D009713:Nucleotidyltransferases / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D001704:Biopolymers; D000242:Cyclic AMP; D009713:Nucleotidyltransferases; D027081:DNA Gyrase
keywords: AMPylation; adenylylation; enzyme regulation; molecular timer; posttranslational modification