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: 9361724
• 负责人: Yue Zou
• 金额: $ 32.85万
• 依托单位: EAST TENNESSEE STATE UNIVERSITY
• 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 2022-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9361724
• 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损伤是人类癌和许多其他人类疾病的主要原因。响应 DNA损伤，细胞激活DNA损伤响应（DDR）途径，例如DNA损伤检查点， DNA修复和凋亡。 ATR（共济失调的Telangiectia and rad3相关），一个成员 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 通过信号DNA损伤，激活检查点，在DDR中起关键作用的蛋白激酶， 阻止细胞周期进程并支撑DNA修复以恢复DNA完整性。有趣的是，一个身体 小鼠模型和人类流行病学研究的证据表明，与ATM不同，密切相关 ATR样的Pikk家族成员，其缺乏促进癌变，ATR​​抑制作用抑制 致癌作用。此外，ATR淘汰赛在胚胎上是致命的。这些暗示了ATR的参与 在调节细胞死亡时。尽管ATR已广泛研究了DDR的检查点激酶 核，对其在细胞质或线粒体（细胞细胞器）中的功能知之甚少 激活DNA损伤引起的凋亡。 P.I.实验室的最新发现表明，（a）除了它 Hallmark核检查点函数，ATR是直接在线粒体上运作的生存蛋白 反对紫外线伤害； （b）ATR包含一个BH3样结构域，使ATR像Bcl-2家族蛋白一样起作用； （c）重要的是，线粒体ATR是一种由PIN1调节的ATR的Prolyl cis-异构形式，而相反， 核ATR是ATR的跨异构体形式；最后（d）ATR的线粒体活性独立于 它的检查点激酶活性和ATTRIP。在这个项目中，我们将测试（1）prolyl的假设 异构化改变ATR的结构，将ATR功能转化为线粒体特异性活动 促进细胞存活或核功能作为DNA损伤检查点调节剂； （2）抗凋亡活性 线粒体的ATR在体内介导癌变中起重要作用，因此抑制 这种活动可能会减少癌变/肿瘤发生，并为预防癌症和 治疗; （3）ATR的DNA损伤检查点活动需要ATR的反异构形式，并在 ATR和/或ATR-ATRIP复合物的翻译修饰可能在稳定ATR中发挥作用 在细胞核中的反异构形式中。这些假设将在以下特定目标中进行检验。 目的1：定义ATR丙酰异构体的结构功能关系，ATRH-TBID和ATRH- PIN1相互作用；目的2：确定脯氨酰同源化在ATR核功能中的作用；和 目标3：确定ATR异构体对癌变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，也有 对其他人类疾病（例如神经退行性和心血管疾病）的影响。