genetic screen /lab/aaron-whiteley/ en Analysis of human cGAS activity and structure /lab/aaron-whiteley/2019/05/02/analysis-human-cgas-activity-and-structure Analysis of human cGAS activity and structure Anonymous (not verified) Thu, 05/02/2019 - 00:00 DNA sensing STING X-ray crystallography biochemistry cGAS genetic screen innate immunity nucleotidyltransferase Zhou W ,  ➤Whiteley AT ,  Kranzusch PJ

Methods Enzymol. 2019;625:13-40. doi: 10.1016/bs.mie.2019.04.012. Epub 2019 May 2.

Abstract

Cyclic GMP-AMP synthase (cGAS) is an innate immune system enzyme responsible for recognition of double-stranded DNA aberrantly localized in the cell cytosol. cGAS binds DNA and is activated to catalyze production of the nucleotide second messenger 2'-5'/3'-5' cyclic GMP-AMP (2'3' cGAMP). In spite of a major role for cGAS in the cellular immune response, a complete understanding of cGAS biology has been limited by a lack of genetic tools to rapidly screen cGAS activity, instability of human cGAS-DNA interactions in vitro, and a previous absence of structural information for the human cGAS-DNA complex. Here we detail procedures to map the molecular determinants of cGAS activation and describe methods developed to prepare human cGAS-DNA crystals for structural analysis. Together with earlier systems established to study mammalian homologs of cGAS, these innovations provide a foundation to understand and therapeutically target human cGAS biology.

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Zhou W, ➤Whiteley AT, Kranzusch PJ. | Methods Enzymol. 2019

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