title: The cyclic dinucleotide c-di-AMP is an allosteric regulator of metabolic enzyme function.
abstract: Cyclic di-adenosine monophosphate (c-di-AMP) is a broadly conserved second messenger required for bacterial growth and infection. However, the molecular mechanisms of c-di-AMP signaling are still poorly understood. Using a chemical proteomics screen for c-di-AMP-interacting proteins in the pathogen Listeria monocytogenes, we identified several broadly conserved protein receptors, including the central metabolic enzyme pyruvate carboxylase (LmPC). Biochemical and crystallographic studies of the LmPC-c-di-AMP interaction revealed a previously unrecognized allosteric regulatory site 25 Å from the active site. Mutations in this site disrupted c-di-AMP binding and affected catalytic activity of LmPC as well as PC from pathogenic Enterococcus faecalis. C-di-AMP depletion resulted in altered metabolic activity in L. monocytogenes. Correction of this metabolic imbalance rescued bacterial growth, reduced bacterial lysis, and resulted in enhanced bacterial burdens during infection. These findings greatly expand the c-di-AMP signaling repertoire and reveal a central metabolic regulatory role for a cyclic dinucleotide.
authors.affiliation: Department of Microbiology, University of Washington, Seattle, WA 98195, USA.
authors.forename: Kamakshi
authors.initials: K
authors.lastname: Sureka
authors.affiliation: Department of Biological Sciences, Columbia University, New York, NY 10027, USA.
authors.forename: Philip H
authors.initials: PH
authors.lastname: Choi
authors.affiliation: Department of Microbiology, University of Washington, Seattle, WA 98195, USA.
authors.forename: Mimi
authors.initials: M
authors.lastname: Precit
authors.affiliation: Department of Molecular and Cell Biology, University of California, Berkeley, CA 94720, USA.
authors.forename: Matthieu
authors.initials: M
authors.lastname: Delince
authors.affiliation: Department of Medical Microbiology and Immunology, University of Wisconsin-Madison, Madison, WI 53706, USA.
authors.forename: Daniel A
authors.initials: DA
authors.lastname: Pensinger
authors.affiliation: Department of Microbiology, University of Washington, Seattle, WA 98195, USA.
authors.forename: TuAnh Ngoc
authors.initials: TN
authors.lastname: Huynh
authors.affiliation: Department of Biological Sciences, Columbia University, New York, NY 10027, USA.
authors.forename: Ashley R
authors.initials: AR
authors.lastname: Jurado
authors.affiliation: Department of Pharmaceutical Sciences, University of Maryland, Baltimore, MD 21201, USA.
authors.forename: Young Ah
authors.initials: YA
authors.lastname: Goo
authors.affiliation: Department of Chemistry, University of Washington, Seattle, WA 98195, USA.
authors.forename: Martin
authors.initials: M
authors.lastname: Sadilek
authors.affiliation: QB3/Chemistry Mass Spectrometry Facility, University of California, Berkeley, CA 94720, USA.
authors.forename: Anthony T
authors.initials: AT
authors.lastname: Iavarone
authors.affiliation: Department of Medical Microbiology and Immunology, University of Wisconsin-Madison, Madison, WI 53706, USA.
authors.forename: John-Demian
authors.initials: JD
authors.lastname: Sauer
authors.affiliation: Department of Biological Sciences, Columbia University, New York, NY 10027, USA. Electronic address: ltong@columbia.edu.
authors.forename: Liang
authors.initials: L
authors.lastname: Tong
authors.affiliation: Department of Microbiology, University of Washington, Seattle, WA 98195, USA. Electronic address: jjwoodwa@uw.edu.
authors.forename: Joshua J
authors.initials: JJ
authors.lastname: Woodward
mesh_terms: D000494:Allosteric Regulation; D000595:Amino Acid Sequence; D000818:Animals; D001433:Bacteriolysis; D001665:Binding Sites; D018360:Crystallography, X-Ray; D015226:Dinucleoside Phosphates / Q000378:metabolism*; D054884:Host-Pathogen Interactions; D008089:Listeria monocytogenes / Q000201:enzymology / Q000254:growth & development / Q000378:metabolism*; D008088:Listeriosis / Q000382:microbiology; D051379:Mice; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011766:Pyruvate Carboxylase / Q000737:chemistry* / Q000378:metabolism*
chemical_list: D015226:Dinucleoside Phosphates; C528998:cyclic diadenosine phosphate; D011766:Pyruvate Carboxylase