ATR Isomerization in Cellular Responses to UV Damage of DNA

细胞对 DNA 紫外线损伤反应中的 ATR 异构化

• 批准号: 10220898
• 负责人: Yue Zou
• 金额: $ 35.34万
• 依托单位: UNIVERSITY OF TOLEDO HEALTH SCI CAMPUS
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-08-01 至 2023-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10220898
• 关键词: ATR gene; Address; Adopted; Aging; Amino Acids; Apoptosis; Apoptotic; BCL2 gene; Cancer Biology; Cardiovascular Diseases; Cell Cycle Progression; Cell Death; Cell Nucleus; Cell Survival; Cells; Clinic; Clinical Trials; Complex; Cytoplasm; DNA; DNA Damage; DNA Repair; DNA damage checkpoint; Embryo; Exhibits; Family; Family member; Human; Isomerism; KRP protein; Knock-out; Laboratories; Malignant Neoplasms; Malignant neoplasm of ovary; Maps; Mass Spectrum Analysis; Mediating; Mitochondria; Molecular Conformation; Mutant Strains Mice; Mutate; NIMA; Neurodegenerative Disorders; Nuclear; Organelles; Pathway interactions; Phosphatidylinositols; Phosphorylation; Phosphotransferases; Play; Post-Translational Protein Processing; Protein Dephosphorylation; Protein Family; Protein Footprinting; Protein Kinase; Proteins; Role; Signal Transduction; Structure; Structure-Activity Relationship; Study models; TREX1 gene; Telangiectasis; Testing; UV carcinogenesis; UV induced; UV induced DNA damage; ataxia telangiectasia mutated protein; base; cancer prevention; cancer therapy; carcinogenesis; cis trans isomerization; cis-trans-Isomerases; cytochrome c; epidemiology study; genome integrity; human disease; in vivo; innovation; member; mouse model; novel; novel strategies; protein function; response; therapeutic target; tumorigenesis; ultraviolet damage

Summary DNA damage is a major cause of human cancers and many other human diseases. In response to DNA damage, cells activate DNA damage response (DDR) pathways such as DNA damage checkpoints, DNA repair, and apoptosis. ATR (ataxia telangiectasia and Rad3-related), a member of the phosphoinositide 3-kinase-related protein kinases (PIKK) family, is a major DNA damage checkpoint protein kinase which plays a critical role in DDR by signaling DNA damage, activating checkpoints, arresting cell cycle progression and facilitating DNA repair to restore DNA integrity. Interestingly, a body of evidence from mouse model and human epidemiologic studies shows that unlike ATM, a closely related ATR-like PIKK family member whose deficiency promotes carcinogenesis, ATR inhibition suppresses carcinogenesis. Moreover, ATR knockout is embryonically lethal. These suggest an involvement of ATR in regulating cell death. Although ATR has been extensively studied as a checkpoint kinase in DDR in the nucleus, little is known about its functions in the cytoplasm or mitochondria, the cellular organelle for activating DNA damage-induced apoptosis. A recent finding from the P.I.’s lab reveals that (a) besides its hallmark nuclear checkpoint functions, ATR is a pro-survival protein functioning directly at mitochondria against UV damage; (b) ATR contains a BH3-like domain that allows ATR to act like a Bcl-2 family protein; (c) importantly, mitochondrial ATR is a prolyl cis-isomeric form of ATR regulated by Pin1 while in contrast, nuclear ATR is a trans-isomeric form of ATR; and finally (d) mitochondria activity of ATR is independent of its checkpoint kinase activity and ATRIP. In this project, we will test the hypotheses that (1) Prolyl isomerization alters the structure of ATR, transforming ATR functions for mitochondria-specific activities to promote cell survival or nuclear functions as a DNA damage checkpoint regulator; (2) Antiapoptotic activity of ATR at mitochondria plays an important role in mediating carcinogenesis in vivo, and thus, suppressing such activity may reduce carcinogenesis/tumorigenesis and provide a strategy for cancer prevention and treatment; and (3) ATR’s trans-isomeric form is required for its DNA damage checkpoint activity, and post- translational modifications of ATR and/or ATR-ATRIP complex formation may play a role in stabilizing ATR in the trans-isomeric form in the nucleus. These hypotheses will be tested in the following specific aims. Aim 1: To define the structure-function relationships of ATR prolyl isomers, and of ATRH-tBid and ATRH- Pin1 interactions; Aim 2: To determine the role of prolyl isomerization in the nuclear functions of ATR; and Aim 3: To determine the in vivo effects of ATR isomers on carcinogenesis and tumorigenesis. The proposed studies represent an innovative effort highly relevant to cancer biology and also having implications in other human diseases such as neurodegenerative and cardiovascular diseases.

概括 DNA 损伤是人类癌症和许多其他人类疾病的主要原因。作为回应 DNA损伤，细胞激活DNA损伤反应（DDR）途径，例如DNA损伤检查点， DNA 修复和细胞凋亡。 ATR（毛细血管扩张性共济失调和 Rad3 相关）， 磷酸肌醇 3 激酶相关蛋白激酶 (PIKK) 家族是主要的 DNA 损伤检查点 蛋白激酶通过发出 DNA 损伤信号、激活检查点，在 DDR 中发挥关键作用， 阻止细胞周期进程并促进 DNA 修复以恢复 DNA 完整性。有趣的是，一个身体 来自小鼠模型和人类流行病学研究的证据表明，与 ATM 不同，一种密切相关的 ATR 样 PIKK 家族成员，其缺陷促进致癌，ATR 抑制抑制 致癌作用。此外，ATR 敲除具有胚胎致死性。这些表明 ATR 的参与 调节细胞死亡。尽管 ATR 作为 DDR 中的检查点激酶已被广泛研究 细胞核，但对其在细胞质或线粒体（细胞器）中的功能知之甚少。 激活DNA损伤诱导的细胞凋亡。 P.I. 实验室最近的一项发现表明，(a) 除了 ATR 是一种标志性核检查点功能，是一种直接在线粒体上发挥作用的促生存蛋白 防止紫外线伤害； (b) ATR 含有 BH3 样结构域，使 ATR 能够像 Bcl-2 家族蛋白一样发挥作用； (c) 重要的是，线粒体 ATR 是 ATR 的脯氨酰顺式异构形式，受 Pin1 调节，而相比之下， 核ATR是ATR的反式异构形式；最后 (d) ATR 的线粒体活性独立于 其检查点激酶活性和 ATRIP。在这个项目中，我们将测试以下假设：(1) Prolyl 异构化改变了 ATR 的结构，将线粒体特异性活动的 ATR 功能转变为 作为 DNA 损伤检查点调节剂促进细胞存活或核功能； (2) 抗凋亡活性 线粒体上的 ATR 在介导体内致癌过程中发挥重要作用，从而抑制 这种活性可能会减少癌变/肿瘤发生，并为癌症预防和治疗提供策略。 治疗; (3) ATR 的反式异构体形式是其 DNA 损伤检查点活性所必需的，并且后 ATR 和/或 ATR-ATRIP 复合物形成的翻译修饰可能在稳定 ATR 中发挥作用 在细胞核中以反式异构体形式存在。这些假设将在以下具体目标中得到检验。 目标 1：定义 ATR 脯氨酰异构体、ATRH-tBid 和 ATRH- 的结构-功能关系 Pin1 交互；目标2：确定脯氨酰异构化在ATR核功能中的作用；和 目标 3：确定 ATR 异构体对癌发生和肿瘤发生的体内影响。这 拟议的研究代表了与癌症生物学高度相关的创新努力，并且还具有 对神经退行性疾病和心血管疾病等其他人类疾病的影响。