Non-canonical cGAS signaling in DNA damage response

DNA 损伤反应中的非典型 cGAS 信号传导

基本信息

批准号：
10307611
负责人：
Nagaraj Kerur
金额：
$ 48.63万
依托单位：
OHIO STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-12-05 至 2025-11-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10307611
关键词：
ATM activation Address Adenosine Monophosphate Affinity Aging Apical BRCA1 gene Binding Biochemical Biological Process CHEK1 gene CHEK2 gene CRISPR/Cas technology Cell Aging Cell Cycle Cell Cycle Arrest Cell Cycle Checkpoint Cell Death Cells Chemical Agents Chromatin Complex Cyclic GMP Cytosol DNA DNA Damage DNA Double Strand Break DNA Repair DNA biosynthesis DNA strand break Data Defense Mechanisms Dimerization Dinucleoside Phosphates Disulfides Embryo Endoplasmic Reticulum Enzymes Eukaryotic Cell Exposure to Foundations G1 Arrest Generations Genes Genetic Transcription Genome Genomic DNA Genomic Instability Guanosine Triphosphate HTATIP gene Histones Homeostasis Human IFNAR1 gene IRF3 gene Immune system Immunologic Receptors Inflammatory Response Interferon Type I Interferons Invertebrates Laboratories Left Lymphocyte Maintenance Malignant Neoplasms Manuscripts Mediating Mediator Membrane Proteins Mitotic Molecular Mus Natural Immunity Neoplasm Metastasis Oxidative Stress Oysters Pathogen detection Pathway interactions Pattern recognition receptor Periodicity Process Role STAT2 gene Sea Anemones Second Messenger Systems Signal Pathway Signal Transduction Stimulus System TANK-binding kinase 1 TP53 gene Telomerase V(D)J Recombination Viral Work assault ataxia telangiectasia mutated protein crosslink cytokine endodeoxyribonuclease SceI endonuclease genome integrity immune function insight micronucleus novel pathogen receptor recruit repaired response sensor tumorigenesis

项目摘要

SUMMARY The vertebrate innate immunity relies upon a complex set of cytosolic pattern recognition receptors (PRR) to detect pathogen-derived, evolutionarily conserved molecules such as DNA. The sensing of cytosolic DNA by cyclic-GMP-AMP synthase (cGAS) activates the enzymatic synthesis of cyclic guanosine monophosphate- adenosine monophosphate (cGAMP), a cyclic dinucleotide (CDN) second messenger. cGAMP signals via its high affinity receptor protein STING, which subsequently recruits TANK-binding kinase 1 (TBK1) and interferon (IFN) regulatory factor 3 (IRF3) to stimulate the induction of type I IFNs. Although originally identified as a cytosolic sensor of foreign DNA, cGAS is also recruited to and activated by fragments of chromatin from damaged genomic DNA in the cytosol or micronuclei. Multiple convergent studies have recently highlighted the significance of cGAS in the DNA damage–induced inflammatory response and its implications for cellular senescence, tumorigenesis, and metastasis. Nothing, however, is known about whether cGAS activation in these contexts directly contributes to the maintenance of genome integrity. Recent studies in our laboratory have discovered that cGAS/cGAMP signaling triggers DNA damage response (DDR), independently of its well-characterized type I interferon pathway. These studies revealed that cGAMP-induced DDR activates cell cycle checkpoint responses that lead to G1 arrest and subsequent suppression of homology directed repair (HDR) of double-strand DNA breaks (DSB) in CRISPR/Cas9-edited mouse embryos and human and mouse cells. Interestingly, the cGAMP-induced DDR was also demonstrable in invertebrate species (oysters and starlet sea anemone) lacking interferon-based immune system, suggesting that the DNA damage surveillance mechanism of cGAMP predates its more well- known IFN-mediated immune function. The studies proposed here aim to advance these novel findings by elucidating the molecular mechanism of cGAS/cGAMP-induced DDR induction via three thematically integrated, yet independent Aims: (1) Decipher the critical signaling pathways involved in cGAMP-induced ATM activation; (2) Define the molecular mechanism of cGAS-cGAMP-induced suppression of HDR; and (3) Elucidate whether cGAMP-induced DDR potentiates the cGAS-initiated innate immunity. In summary, this work will illuminate novel aspects of the molecular and biochemical basis of cGAMP-induced activation of the apical DDR signaling kinase ATM, and increase understanding of the relationship between cGAMP-induced DDR signaling and the traditional immune function of cGAS.

概括脊椎动物的先天免疫依赖于一组复杂的胞质模式识别受体（PRR）来检测病原体衍生的、进化上保守的分子，例如 DNA 通过感测胞质 DNA。环-GMP-AMP 合酶 (cGAS) 激活环单磷酸鸟苷的酶促合成单磷酸腺苷 (cGAMP)，一种环状二核苷酸 (CDN) 第二信使，通过其 cGAMP 发出信号。高亲和力受体蛋白 STING，随后招募 TANK 结合激酶 1 (TBK1) 和干扰素 (IFN) 调节因子 3 (IRF3) 刺激 I 型 IFN 的诱导，尽管最初被鉴定为一种。作为外源 DNA 的胞质传感器，cGAS 也被来自染色质片段的募集并激活。细胞质或微核中的基因组 DNA 受损。最近多项趋同研究强调了 cGAS 在 DNA 损伤诱导中的重要性炎症反应及其对细胞衰老、肿瘤发生和转移的影响。然而，我们知道在这些情况下 cGAS 激活是否直接有助于维持我们实验室最近的研究发现 cGAS/cGAMP 信号触发。 DNA 损伤反应 (DDR)，独立于其特征明确的 I 型干扰素途径。研究表明，cGAMP 诱导的 DDR 激活细胞周期检查点反应，导致 G1 期停滞以及随后抑制双链 DNA 断裂 (DSB) 的同源定向修复 (HDR) CRISPR/Cas9 编辑的小鼠胚胎以及人类和小鼠细胞。在缺乏干扰素的无脊椎动物（牡蛎和小星海葵）中也得到了证实免疫系统，这表明 cGAMP 的 DNA 损伤监视机制早于它的更完善。已知IFN介导的免疫功能。这里提出的研究旨在通过阐明其分子机制来推进这些新发现 cGAS/cGAMP 通过三个主题整合但独立的 DDR 诱导目标：(1) 解密 (2) 定义分子 cGAS-cGAMP 诱导的 HDR 抑制机制；以及 (3) 阐明 cGAMP 是否诱导 DDR 增强 cGAS 引发的先天免疫总而言之，这项工作将阐明 cGAS 的新颖方面。 cGAMP 诱导顶端 DDR 信号激酶 ATM 激活的分子和生化基础，以及增加对 cGAMP 诱导的 DDR 信号传导与传统免疫之间关系的了解 cGAS 的功能。