# Molecular basis of viral DNA sensing through the cGAS-STING pathway

> **NIH NIH R01** · TEXAS A&M AGRILIFE RESEARCH · 2020 · $362,292

## Abstract

The Molecular Basis of Viral DNA Sensing through the cGAS-STING Pathway
The innate immune response is the first line of defense against invading pathogens. Viral nucleic acids, such
as DNA and RNA, serve as danger signals that induce potent immune responses in infected cells. The
detection of viral nucleic acids is a central strategy by which the hosts sense infection and launch protective
immune responses. Cyclic GMP-AMP synthase (cGAS) is a cytosolic dsDNA sensor in innate immunity. It is
activated by viral DNA and catalyzes the synthesis of a noncanonical cyclic dinucleotide 2ʹ, 3ʹ cGAMP (referred
to as cGAMP). As a second messenger, cGAMP stimulates the induction of type I interferons (IFNs) via the
adaptor STING located on the ER membrane. The engagement of cGAMP by STING leads to the recruitment
and activation of the protein kinase TBK1. The transcriptional factor IRF-3 is then recruited to the signaling
complex by phosphorylated STING. The induced proximity of IRF-3 and TBK1 leads to IRF-3 phosphorylation
and activation. Phosphorylated IRF-3 oligomerizes, translocates to the nucleus, and induces the expression of
type I IFNs, conferring antiviral activity to the host and activating the acquired immune responses. To
understand the molecular basis of viral DNA sensing through the cGAS-STING pathway, the PI determined the
crystal structures of cGAS in isolation and in complex with dsDNA; the structures of STING in isolation and in
complex with cGAMP; the structures of mouse TBK1 bound to two inhibitors; the structure of phosphorylated
STING C-terminal peptide bound to IRF-3; and the structure of a phosphomimetic dimer of IRF-3. These
comprehensive structural and functional studies significantly advanced our understanding about how the innate
immune system responds to viral DNA at molecular level. However, it remains unknown how cGAMP binding
activates STING and mediates the recruitment and activation of TBK1 and IRF-3 at the signaling complex. In
addition, the exact mechanism of how phosphorylation induces the activation of IRF-3 and facilitates its
association with the IFN-β promoter remains to be determined. Built-upon our strong preliminary studies, the
proposed research will elucidate the molecular bases of several key steps of the cGAS-STING pathway with
the following aims: 1). Determine the structural basis of TBK1 recruitment by STING; 2). Elucidate the
molecular basis of STING activation by cGAMP; 3). Delineate the mechanism of IRF-3 activation by TBK1. The
proposed studies represent a rigorous and comprehensive investigation into the mechanisms of cytosolic DNA
sensing and will dramatically advance our understanding of the molecular basis of innate immunity against viral
infection. In addition, they provide a structural framework for innovative approaches to treat viral diseases,
autoimmune disorders, and cancer by manipulating the innate immune response.

## Key facts

- **NIH application ID:** 9875428
- **Project number:** 5R01AI145287-02
- **Recipient organization:** TEXAS A&M AGRILIFE RESEARCH
- **Principal Investigator:** Pingwei Li
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $362,292
- **Award type:** 5
- **Project period:** 2019-02-20 → 2024-01-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/9875428

## Citation

> US National Institutes of Health, RePORTER application 9875428, Molecular basis of viral DNA sensing through the cGAS-STING pathway (5R01AI145287-02). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/9875428. Licensed CC0.

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