title: Identification and characterization of phosphodiesterases that specifically degrade 3'3'-cyclic GMP-AMP.
abstract: Cyclic dinucleotides act as intracellular second messengers, modulating a variety of cellular activities including innate immune activation. Although phosphodiesterases (PDEs) hydrolyzing c-di-GMP and c-di-AMP have been identified, no PDEs for cGAMPs have been reported. Here we identified the first three cGAMP-specific PDEs in V. cholerae (herein designated as V-cGAP1/2/3). V-cGAPs are HD-GYP domain-containing proteins and specifically break 3'3'-cGAMP, but not other forms of cGAMP. 3'3'-cGAMP is first linearized by all three V-cGAPs to produce 5'-pApG, which is further hydrolyzed into 5'-ApG by V-cGAP1. In this two-step reaction, V-cGAP1 functions as both a PDE and a 5'-nucleotidase. In vivo experiments demonstrated that V-cGAPs play non-redundant roles in cGAMP degradation. The high specificity of V-cGAPs on 3'3'-cGAMP suggests the existence of specific PDEs for other cGAMPs, including 2'3'-cGAMP in mammalian cells. The absolute requirement of the GYP motif for 3'3'-cGAMP degradation suggests that HD domain-containing PDEs in eukaryotes are probably unable to hydrolyze cGAMPs. The fact that all V-cGAPs attack 3'3'-cGAMP on one specific phosphodiester bond suggests that PDEs for other cGAMPs would utilize a similar strategy. These results will provide valuable information for identification and characterization of mammalian 2'3'-cGAMP-specific PDEs in future studies.
authors.affiliation: 1] State Key Laboratory of Protein and Plant Gene Research, School of Life Sciences, Peking University, Beijing 100871, China [2] Key Laboratory of Cell Proliferation and Differentiation of the Ministry of Education, School of Life Sciences, Peking University, Beijing 100871, China [3] Peking-Tsinghua Center for Life Sciences, Beijing, China.
authors.forename: Juyi
authors.initials: J
authors.lastname: Gao
authors.affiliation: 1] State Key Laboratory of Protein and Plant Gene Research, School of Life Sciences, Peking University, Beijing 100871, China [2] Key Laboratory of Cell Proliferation and Differentiation of the Ministry of Education, School of Life Sciences, Peking University, Beijing 100871, China [3] Peking-Tsinghua Center for Life Sciences, Beijing, China.
authors.forename: Jianli
authors.initials: J
authors.lastname: Tao
authors.affiliation: 1] State Key Laboratory for Infectious Disease Prevention and Control, National Institute for Communicable Disease Control and Prevention, Chinese Centre for Disease Control and Prevention, Beijing 102206, China [2] Collaborative Innovation Centre for Diagnosis and Treatment of Infectious Diseases, Hangzhou 310003, China.
authors.forename: Weili
authors.initials: W
authors.lastname: Liang
authors.affiliation: State Key Laboratory for Infectious Disease Prevention and Control, National Institute for Communicable Disease Control and Prevention, Chinese Centre for Disease Control and Prevention, Beijing 102206, China.
authors.forename: Meng
authors.initials: M
authors.lastname: Zhao
authors.affiliation: State Key Laboratory of Protein and Plant Gene Research, School of Life Sciences, Peking University, Beijing 100871, China.
authors.forename: Xiaoxia
authors.initials: X
authors.lastname: Du
authors.affiliation: Biodynamic Optical Imaging Center (BIOPIC), School of Life Sciences, Peking University, Beijing 100871, China.
authors.forename: Shan
authors.initials: S
authors.lastname: Cui
authors.affiliation: Biodynamic Optical Imaging Center (BIOPIC), School of Life Sciences, Peking University, Beijing 100871, China.
authors.forename: Haifeng
authors.initials: H
authors.lastname: Duan
authors.affiliation: 1] State Key Laboratory for Infectious Disease Prevention and Control, National Institute for Communicable Disease Control and Prevention, Chinese Centre for Disease Control and Prevention, Beijing 102206, China [2] Collaborative Innovation Centre for Diagnosis and Treatment of Infectious Diseases, Hangzhou 310003, China.
authors.forename: Biao
authors.initials: B
authors.lastname: Kan
authors.affiliation: 1] State Key Laboratory of Protein and Plant Gene Research, School of Life Sciences, Peking University, Beijing 100871, China [2] Biodynamic Optical Imaging Center (BIOPIC), School of Life Sciences, Peking University, Beijing 100871, China.
authors.forename: Xiaodong
authors.initials: X
authors.lastname: Su
authors.affiliation: 1] State Key Laboratory of Protein and Plant Gene Research, School of Life Sciences, Peking University, Beijing 100871, China [2] Key Laboratory of Cell Proliferation and Differentiation of the Ministry of Education, School of Life Sciences, Peking University, Beijing 100871, China [3] Peking-Tsinghua Center for Life Sciences, Beijing, China.
authors.forename: Zhengfan
authors.initials: Z
authors.lastname: Jiang
mesh_terms: D000818:Animals; D018441:Biofilms / Q000254:growth & development; D002851:Chromatography, High Pressure Liquid; D015226:Dinucleoside Phosphates / Q000378:metabolism; D051379:Mice; D009712:Nucleotides, Cyclic / Q000378:metabolism; D010727:Phosphoric Diester Hydrolases / Q000235:genetics / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D014734:Vibrio cholerae / Q000201:enzymology / Q000254:growth & development
chemical_list: D015226:Dinucleoside Phosphates; D009712:Nucleotides, Cyclic; C528998:cyclic diadenosine phosphate; C584311:cyclic guanosine monophosphate-adenosine monophosphate; D010727:Phosphoric Diester Hydrolases
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