@article{568b5a5333d745f1a50a042f5841fa01,
title = "Structure-guided reprogramming of human cgas dinucleotide linkage specificity",
abstract = "Cyclic dinucleotides (CDNs) play central roles in bacterial pathogenesis and innate immunity. The mammalian enzyme cGAS synthesizes a unique cyclic dinucleotide (cGAMP) containing a 2Ephosphodiester linkage essential for optimal immune stimulation, but the molecular basis for linkage specificity is unknown. Here, we show that the Vibrio cholerae pathogenicity factor DncV is a prokaryotic cGAS-like enzyme whose activity provides a mechanistic rationale for the unique ability of cGAS to produce 2E-5E cGAMP. Three high-resolution crystal structures show that DncV and human cGAS generate CDNs in sequential reactions that proceed in opposing directions. We explain 2{\^E}and 3{\^E} linkage specificity and test this model by reprogramming the human cGAS active site to produce 3{\^E}-5{\^E} cGAMP, leading to selective stimulation of alternative STING adaptor alleles in cells. These results demonstrate mechanistic homology between bacterial signaling and mammalian innate immunity and explain how active site configuration controls linkage chemistry for pathway-specific signaling.",
author = "Kranzusch, {Philip J.} and Lee, {Amy S.Y.} and Wilson, {Stephen C.} and Solovykh, {Mikhail S.} and Vance, {Russell E.} and Berger, {James M.} and Doudna, {Jennifer A.}",
note = "Funding Information: X-ray data were collected at Beamline 8.3.1 of the Lawrence Berkeley National Lab Advanced Light Source (ALS) and Beamlines 11.1 and 12.2 of the Stanford Synchrotron Radiation Lightsource (SSRL). The authors are grateful to J. Holton and G. Meigs (ALS); T. Doukov, S. Russi, and A. Gonzalez (SSRL) for technical assistance with data collection and processing; R. Nichiporuk (QB3 Mass Spectrometry Facility) for assistance with product determination; J. Cate, Y. Bai, S. Floor, R. Wilson, and members of the Doudna and Berger labs for helpful comments and discussion. This work was funded by HHMI (R.E.V. and J.A.D.); the G. Harold and Leila Y. Mathers Foundation (J.M.B.); National Cancer Institutes Grant CA077373 (J.M.B.); National Institutes of Health Grants AI063302, AI080749, and AI103817 (R.E.V.); and the NIGMS Center for RNA Systems Biology (A.S.Y.L. and J.A.D.). P.J.K. is supported as an HHMI Fellow of the Life Sciences Research Foundation, and A.S.Y.L. is supported as an American Cancer Society Postdoctoral Fellow (PF-14-108-01-RMC). R.E.V. and J.A.D. are HHMI Investigators. ",
year = "2014",
month = aug,
day = "28",
doi = "10.1016/j.cell.2014.07.028",
language = "English (US)",
volume = "158",
pages = "1011--1021",
journal = "Cell",
issn = "0092-8674",
publisher = "Cell Press",
number = "5",
}