title: Cyclic [G(2',5')pA(3',5')p] is the metazoan second messenger produced by DNA-activated cyclic GMP-AMP synthase.
abstract: Recent studies identified cyclic GMP-AMP (cGAMP) as a metazoan second messenger triggering an interferon response. cGAMP is generated from GTP and ATP by cytoplasmic dsDNA sensor cGAMP synthase (cGAS). We combined structural, chemical, biochemical, and cellular assays to demonstrate that this second messenger contains G(2',5')pA and A(3',5')pG phosphodiester linkages, designated c[G(2',5')pA(3',5')p]. We show that, upon dsDNA binding, cGAS is activated through conformational transitions, resulting in formation of a catalytically competent and accessible nucleotide-binding pocket for generation of c[G(2',5')pA(3',5')p]. We demonstrate that cyclization occurs in a stepwise manner through initial generation of 5'-pppG(2',5')pA prior to cyclization to c[G(2',5')pA(3',5')p], with the latter positioned precisely in the catalytic pocket. Mutants of cGAS dsDNA-binding or catalytic pocket residues exhibit reduced or abrogated activity. Our studies have identified c[G(2',5')pA(3',5')p] as a founding member of a family of metazoan 2',5'-containing cyclic heterodinucleotide second messengers distinct from bacterial 3',5' cyclic dinucleotides.
authors: Structural Biology Program, Memorial Sloan-Kettering Cancer Center, New York, NY 10065, USA.
authors: Pu
authors: P
authors: Gao
authors: Manuel
authors: M
authors: Ascano
authors: Yang
authors: Y
authors: Wu
authors: Winfried
authors: W
authors: Barchet
authors: Barbara L
authors: BL
authors: Gaffney
authors: Thomas
authors: T
authors: Zillinger
authors: Artem A
authors: AA
authors: Serganov
authors: Yizhou
authors: Y
authors: Liu
authors: Roger A
authors: RA
authors: Jones
authors: Gunther
authors: G
authors: Hartmann
authors: Thomas
authors: T
authors: Tuschl
authors: Dinshaw J
authors: DJ
authors: Patel
mesh_terms: D015088:2',5'-Oligoadenylate Synthetase / Q000737:chemistry; D000255:Adenosine Triphosphate / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D018360:Crystallography, X-Ray; D004247:DNA / Q000737:chemistry / Q000378:metabolism; D015226:Dinucleoside Phosphates / Q000378:metabolism*; D006160:Guanosine Triphosphate / Q000378:metabolism; D006801:Humans; D051379:Mice; D008956:Models, Chemical; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009712:Nucleotides, Cyclic / Q000378:metabolism*; D009713:Nucleotidyltransferases / Q000737:chemistry* / Q000378:metabolism; D015290:Second Messenger Systems; D016415:Sequence Alignment; D000099235:Cyclic Guanosine Monophosphate-Adenosine Monophosphate Synthase
chemical_list: D015088:2',5'-Oligoadenylate Synthetase; D000255:Adenosine Triphosphate; D004247:DNA; D015226:Dinucleoside Phosphates; D006160:Guanosine Triphosphate; D009712:Nucleotides, Cyclic; D009713:Nucleotidyltransferases; C475085:OAS1 protein, human; C578390:cGAS protein, human; D000099235:Cyclic Guanosine Monophosphate-Adenosine Monophosphate Synthase; C578391:cGAS protein, mouse
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