title: An HD-domain phosphodiesterase mediates cooperative hydrolysis of c-di-AMP to affect bacterial growth and virulence.
abstract: The nucleotide cyclic di-3',5'- adenosine monophosphate (c-di-AMP) was recently identified as an essential and widespread second messenger in bacterial signaling. Among c-di-AMP-producing bacteria, altered nucleotide levels result in several physiological defects and attenuated virulence. Thus, a detailed molecular understanding of c-di-AMP metabolism is of both fundamental and practical interest. Currently, c-di-AMP degradation is recognized solely among DHH-DHHA1 domain-containing phosphodiesterases. Using chemical proteomics, we identified the Listeria monocytogenes protein PgpH as a molecular target of c-di-AMP. Biochemical and structural studies revealed that the PgpH His-Asp (HD) domain bound c-di-AMP with high affinity and specifically hydrolyzed this nucleotide to 5'-pApA. PgpH hydrolysis activity was inhibited by ppGpp, indicating a cross-talk between c-di-AMP signaling and the stringent response. Genetic analyses supported coordinated regulation of c-di-AMP levels in and out of the host. Intriguingly, a L. monocytogenes mutant that lacks c-di-AMP phosphodiesterases exhibited elevated c-di-AMP levels, hyperinduced a host type-I IFN response, and was significantly attenuated for infection. Furthermore, PgpH homologs, which belong to the 7TMR-HD family, are widespread among hundreds of c-di-AMP synthesizing microorganisms. Thus, PgpH represents a broadly conserved class of c-di-AMP phosphodiesterase with possibly other physiological functions in this crucial signaling network.
authors.affiliation: Department of Microbiology, University of Washington, Seattle, WA 98195;
authors.forename: TuAnh Ngoc
authors.initials: TN
authors.lastname: Huynh
authors.affiliation: Department of Biological Sciences, Columbia University, New York, NY 10027; and.
authors.forename: Shukun
authors.initials: S
authors.lastname: Luo
authors.affiliation: Department of Medical Microbiology and Immunology, University of Wisconsin, Madison, WI 53706.
authors.forename: Daniel
authors.initials: D
authors.lastname: Pensinger
authors.affiliation: Department of Medical Microbiology and Immunology, University of Wisconsin, Madison, WI 53706.
authors.forename: John-Demian
authors.initials: JD
authors.lastname: Sauer
authors.affiliation: Department of Biological Sciences, Columbia University, New York, NY 10027; and jjwoodwa@uw.edu ltong@columbia.edu.
authors.forename: Liang
authors.initials: L
authors.lastname: Tong
authors.affiliation: Department of Microbiology, University of Washington, Seattle, WA 98195; jjwoodwa@uw.edu ltong@columbia.edu.
authors.forename: Joshua J
authors.initials: JJ
authors.lastname: Woodward
mesh_terms: D000595:Amino Acid Sequence; D000242:Cyclic AMP / Q000378:metabolism*; D006868:Hydrolysis; D008089:Listeria monocytogenes / Q000201:enzymology* / Q000254:growth & development / Q000472:pathogenicity; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010727:Phosphoric Diester Hydrolases / Q000737:chemistry / Q000378:metabolism*; D011485:Protein Binding; D014774:Virulence
chemical_list: D000242:Cyclic AMP; D010727:Phosphoric Diester Hydrolases
keywords: HD domain; Listeria monocytogenes; bacterial signal transduction; c-di-AMP; phosphodiesterase