ADP-ribosylation Cycles

ADP-核糖基化循环

基本信息

批准号：
10929075
负责人：
Joel Moss
金额：
$ 138.8万
依托单位：
NATIONAL HEART, LUNG, AND BLOOD INSTITUTE
依托单位国家：
美国
项目类别：
财政年份：
资助国家：
美国
起止时间：
至
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10929075
关键词：
2019-nCoV ADP Ribose Transferases ADP ribosylation Adenosine Diphosphate Ribose Amino Acids Annual Reports Arginine Bacteria Bacterial Toxins Binding CD8-Positive T-Lymphocytes COVID-19 COVID-19 therapeutics Cardiac Catalytic Domain Cell Death Cell Differentiation process Cell Nucleus Cell physiology Cells Cessation of life Cholera Toxin Cytoplasm DNA DNA Repair Deacetylation Diphtheria Toxin Echocardiography Elements Embryo Enzymes Exhibits Family Family member Fibroblasts GTP-Binding Protein alpha Subunits, Gs Generations Glycoside Hydrolases Guanine Nucleotides Heart Heterozygote Histone H3 Homeostasis Hydrolase Hydrolysis Immune response Infiltration Inflammation Inflammatory Injury Knockout Mice Laboratories Link Macrophage Magnetic Resonance Imaging Mammalian Cell Mediating Mono(ADP-Ribose) Transferases Mus Myocardial Myocardial Ischemia Niacinamide Nonstructural Protein Nude Mice O-Acetyl-ADP-Ribose Organ Oxidative Stress Pathway interactions Peptides Poly Adenosine Diphosphate Ribose Poly(ADP-ribose) Polymerases Post-Translational Protein Processing Predisposition Process Property Protein Family Protein Kinase Interaction Proteins RIPK1 gene Reaction Regulation Reperfusion Therapy Serine Signal Transduction Sirtuins Specificity Survival Rate TNF gene Tissues Toxin Tumor Promotion Tumorigenicity Up-Regulation Vibrio cholerae Viral Viral Physiology Virulence Virus Xenograft Model age related apoptosis inducing factor arm biological systems carcinogenesis cytokine endonuclease functional group insight male member neoplastic cell pandemic disease pathogenic virus protein function ribose 1-phosphate therapeutic target tumor tumor microenvironment tumorigenesis tumorigenic

项目摘要

Explanation 1.Prior studies demonstrated that cholera toxin, the product of the bacteria Vibrio cholerae, is an ADP-ribosyltransferase that utilizes NAD as a substrate and catalyzes the hydrolysis to ADP-ribose and nicotinamide, as well as transfer of the ADP-ribose to free arginine and arginine residues in proteins. Others showed that the key modified protein was a guanine nucleotide-binding known as G(alpha)s. Based on our findings, we showed that mammalian tissues possessed NAD:arginine ADP-ribosyltransferases. We then asked why the ADP-ribosylation was not toxic, as is the case with cholera toxin, and proposed that mammalian cells had enzymes that hydrolyzed ADP-ribose-arginine(protein). Thus, arginine-specific mono-ADP-ribosylation is a reversible post-translational modification; arginine-specific, cholera toxin-like mono-ADP-ribosyltransferases (ARTC1s) transfer ADP-ribose from NAD+ to arginine, followed by cleavage of ADP-ribose-(arginine)protein bond by ADP-ribosylarginine hydrolase 1 (ARH1), generating unmodified (arginine)protein. ARTC1 has been shown to increase tumorigenicity that is also increased by Arh1 deficiency. In this study, Artc1-KO and Artc1/Arh1-double-KO mice were generated and their properties compared. Artc1-deficient mice showed decreased spontaneous tumorigenesis and increased age-dependent, multi-organ inflammation with upregulation of pro-inflammatory cytokine TNF(alpha). In a xenograft model using tumorigenic Arh1-KO mouse embryonic fibroblasts (MEFs), tumorigenicity was decreased in Artc1-KO and heterozygous recipient mice, with tumor infiltration by CD8+ T cells and macrophages, leading to necroptosis, suggesting that ARTC1 promotes the tumor microenvironment. Furthermore, Artc1/Arh1-double-KO MEFs showed decreased tumorigenesis in nude mice, thus both tumor cells as well as tumor microenvironment require ARTC1. By echocardiography and MRI, Artc1-KO and heterozygous mice showed male-specific, reduced myocardial contractility. Furthermore, Artc1-KO male hearts exhibited enhanced susceptibility to myocardial ischemia-reperfusion-induced injury with increased receptor-interacting protein kinase 3 (RIP3) protein levels compared to WT mice, suggesting that ARTC1 suppresses necroptosis. Overall survival rate of Artc1-KO was less than their Artc1-WT counterparts, primarily due to enhanced immune response and inflammation. Thus, anti-ARTC1 agents may reduce tumorigenesis but may increase multi-organ inflammation and decrease cardiac contractility. 2.COVID 19-related studies: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is responsible for a global pandemic that resulted in more than 6-million deaths worldwide. The virus encodes several non-structural proteins (Nsps) that contain elements capable of disrupting cellular processes. Among these Nsp proteins, Nsp3 contains macrodomains, e.g., Mac1, Mac2, Mac3, with potential effects on host cells. Mac1 has been shown to increase SARS-CoV-2 virulence and disrupt ADP-ribosylation pathways in mammalian cells. Viral macrodomains have been shown to cleave the ADP-ribose-acceptor bond, generating free ADP-ribose. By this reaction, the macrodomain-containing proteins interfere with ADP-ribose homeostasis in host cells. Here, we examined potential hydrolytic activities of SARS-CoV-2 Mac1, 2, and 3 on substrates containing ADP-ribose. Mac1 cleaved -NAD + , but not -NAD + , consistent with stereospecificity at the C-1 bond. This reaction is similar to those catalyzed by ARH1 and ARH3 as well as other macrodomains. In contrast to ARH1 and ARH3, Mac1 did not require Mg 2+ for optimal activity. Mac1 also hydrolyzed O-acetyl-ADP-ribose and ADP-ribose-1-phosphate, but Mac2 and Mac3 were inactive in these reactions. In addition, Mac1 did not cleave -ADP-ribose-(arginine) and ADP-ribose-(serine)-histone H3 peptide, suggesting that Mac1 hydrolyzes ADP-ribose attached to O- and N-linked functional groups, with specificity at the catalytic site in the ADP-ribose moiety. We conclude that SARS-CoV-2 Mac1 may exert anti-viral activity by reversing host-mediated ADP-ribosylation. New insights on Nsp3 Macrodomain activities may shed light on potential SARS-CoV-2 therapeutic targets.

解释 1.先前的研究表明，霍乱毒素是霍乱弧菌的产物，是一种ADP-核糖基转移酶，以NAD为底物，催化水解为ADP-核糖和烟酰胺，以及将ADP-核糖转移为游离蛋白质中的精氨酸和精氨酸残基。其他研究表明，关键的修饰蛋白是鸟嘌呤核苷酸结合蛋白，称为 G(alpha)s。根据我们的发现，我们表明哺乳动物组织具有 NAD：精氨酸 ADP-核糖基转移酶。然后我们询问为什么 ADP-核糖基化不像霍乱毒素那样具有毒性，并提出哺乳动物细胞具有水解 ADP-核糖-精氨酸（蛋白质）的酶。因此，精氨酸特异性单-ADP-核糖基化是一种可逆的翻译后修饰；精氨酸特异性、霍乱毒素样单 ADP-核糖基转移酶 (ARTC1) 将 ADP-核糖从 NAD+ 转移到精氨酸，然后通过 ADP-核糖精氨酸水解酶 1 (ARH1) 裂解 ADP-核糖-（精氨酸）蛋白键，生成未修饰的（精氨酸）蛋白质。 ARTC1 已被证明会增加致瘤性，Arh1 缺陷也会增加致瘤性。在本研究中，生成了 Artc1-KO 和 Artc1/Arh1-double-KO 小鼠并比较了它们的特性。 Artc1缺陷小鼠表现出自发性肿瘤发生减少，年龄依赖性多器官炎症增加，促炎细胞因子TNFα上调。在使用致瘤 Arh1-KO 小鼠胚胎成纤维细胞 (MEF) 的异种移植模型中，Artc1-KO 和杂合受体小鼠的致瘤性降低，CD8+ T 细胞和巨噬细胞浸润肿瘤，导致坏死性凋亡，表明 ARTC1 促进肿瘤微环境。此外，Artc1/Arh1-双-KO MEF在裸鼠中显示出肿瘤发生减少，因此肿瘤细胞和肿瘤微环境都需要ARTC1。通过超声心动图和 MRI，Artc1-KO 和杂合子小鼠表现出雄性特异性的心肌收缩力降低。此外，与 WT 小鼠相比，Artc1-KO 雄性心脏对心肌缺血再灌注诱导的损伤的敏感性增强，受体相互作用蛋白激酶 3 (RIP3) 蛋白水平增加，表明 ARTC1 抑制坏死性凋亡。 Artc1-KO 的总体存活率低于 Artc1-WT 同类，主要是由于免疫反应和炎症增强。因此，抗ARTC1药物可能会减少肿瘤发生，但可能会增加多器官炎症并降低心肌收缩力。 2.COVID 19 相关研究：严重急性呼吸综合征冠状病毒 2 (SARS-CoV-2) 导致全球大流行，导致全球超过 600 万人死亡。该病毒编码多种非结构蛋白（Nsps），其中含有能够破坏细胞过程的元素。在这些 Nsp 蛋白中，Nsp3 含有宏结构域，例如 Mac1、Mac2、Mac3，对宿主细胞具有潜在影响。 Mac1 已被证明可以增加 SARS-CoV-2 的毒力并破坏哺乳动物细胞中的 ADP-核糖基化途径。病毒大结构域已被证明可以裂解 ADP-核糖-受体键，产生游离的 ADP-核糖。通过该反应，含有大结构域的蛋白质干扰宿主细胞中的 ADP-核糖稳态。在这里，我们检查了 SARS-CoV-2 Mac1、2 和 3 对含有 ADP-核糖的底物的潜在水解活性。 Mac1 裂解 -NAD + ，但不裂解 -NAD + ，这与 C-1 键的立体特异性一致。该反应类似于 ARH1 和 ARH3 以及其他宏结构域催化的反应。与 ARH1 和 ARH3 相比，Mac1 不需要 Mg 2+ 即可获得最佳活性。 Mac1 还水解 O-乙酰基-ADP-核糖和 ADP-核糖-1-磷酸，但 Mac2 和 Mac3 在这些反应中不活跃。此外，Mac1 不会切割 -ADP-核糖-（精氨酸）和 ADP-核糖-（丝氨酸）-组蛋白 H3 肽，这表明 Mac1 可以水解附着在 O-和 N-连接官能团上的 ADP-核糖，并且在ADP-核糖部分的催化位点。我们得出的结论是，SARS-CoV-2 Mac1 可能通过逆转宿主介导的 ADP-核糖基化来发挥抗病毒活性。关于 Nsp3 宏结构域活性的新见解可能有助于揭示潜在的 SARS-CoV-2 治疗靶点。

项目成果

期刊论文数量（42）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Cholix toxin, an eukaryotic elongation factor 2 ADP-ribosyltransferase, interacts with Prohibitins and induces apoptosis with mitochondrial dysfunction in human hepatocytes.

Cholix 毒素是一种真核延伸因子 2 ADP-核糖基转移酶，与抑制素相互作用并诱导人肝细胞凋亡和线粒体功能障碍。

DOI：
发表时间：
2019-08
期刊：
影响因子：
3.4
作者：
Yahiro, Kinnosuke;Ogura, Kohei;Terasaki, Yasuhiro;Satoh, Mamoru;Miyagi, Satoru;Terasaki, Mika;Yamasaki, Eiki;Moss, Joel
通讯作者：
Moss, Joel

Novel subtilase cytotoxin produced by Shiga-toxigenic Escherichia coli induces apoptosis in vero cells via mitochondrial membrane damage.

产志贺毒素大肠杆菌产生的新型枯草杆菌酶细胞毒素通过线粒体膜损伤诱导 vero 细胞凋亡。

DOI：
发表时间：
2009-07
期刊：
影响因子：
3.1
作者：
Matsuura, Gen;Morinaga, Naoko;Yahiro, Kinnosuke;Komine, Reiko;Moss, Joel;Yoshida, Hideo;Noda, Masatoshi
通讯作者：
Noda, Masatoshi

Pleiotropic actions of Helicobacter pylori vacuolating cytotoxin, VacA.

幽门螺杆菌空泡细胞毒素 VacA 的多效作用。

DOI：
发表时间：
2010-01
期刊：
影响因子：
0
作者：
Isomoto, Hajime;Moss, Joel;Hirayama, Toshiya
通讯作者：
Hirayama, Toshiya

Identification of subtilase cytotoxin (SubAB) receptors whose signaling, in association with SubAB-induced BiP cleavage, is responsible for apoptosis in HeLa cells.

枯草杆菌酶细胞毒素 (SubAB) 受体的鉴定，其信号传导与 SubAB 诱导的 BiP 裂解相关，负责 HeLa 细胞的凋亡。

DOI：
发表时间：
2011-02
期刊：
影响因子：
3.1
作者：
Yahiro, Kinnosuke;Satoh, Mamoru;Morinaga, Naoko;Tsutsuki, Hiroyasu;Ogura, Kohei;Nagasawa, Sayaka;Nomura, Fumio;Moss, Joel;Noda, Masatoshi
通讯作者：
Noda, Masatoshi

Characterization of Cholix toxin-induced apoptosis in HeLa cells.

Cholix 毒素诱导 HeLa 细胞凋亡的表征。

DOI：
发表时间：
2011-10-28
期刊：
影响因子：
0
作者：
Ogura, Kohei;Yahiro, Kinnosuke;Tsutsuki, Hiroyasu;Nagasawa, Sayaka;Yamasaki, Shinji;Moss, Joel;Noda, Masatoshi
通讯作者：
Noda, Masatoshi