Role of Ubiquitin in Cardiovascular System

泛素在心血管系统中的作用

基本信息

批准号：
8828754
负责人：
YONG TAE KWON
金额：
$ 36.62万
依托单位：
UNIVERSITY OF PITTSBURGH AT PITTSBURGH
依托单位国家：
美国
项目类别：
财政年份：
2006
资助国家：
美国
起止时间：
2006-07-15 至 2017-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8828754
关键词：
Accounting Affinity Amino Acids Aminopeptidase Arginine Arginine Specific tRNA Aspartate Binding Biochemical Biological Availability Blood Blood Vessels Boxing Brain Cardiac Cardiac development Cardiovascular system Cell physiology Cells Cerebrovascular Disorders Cessation of life Chronic Obstructive Airway Disease Complementary DNA Coupled Cysteine Defect Development Dose Drug Targeting Embryo Engineering Environment Family G Protein-Coupled Receptor Genes GTP-Binding Proteins GTPase-Activating Proteins Gene Expression Genes Glutamates Goals Growth Guanosine Triphosphate Half-Life Heart Heart Diseases Homeostasis Homologous Gene Knockout Mice Lead Libraries Licensing Life Link Malignant Neoplasms Mediating Modeling Modification Molecular Morphogenesis Mus Mutant Strains Mice Myocardial N-terminal Names Nature Nitric Oxide Oxygen Pathway interactions Peptides Physiological Play Polyubiquitination Process Protein Biosynthesis Protein Isoforms Proteins Proteolysis Proteomics RGS Proteins Role Signal Transduction Small Interfering RNA Sulfonic Acids Testing Time Tissues Transferase Transgenic Mice Ubiquitin Ubiquitination Ventricular Ventricular Septal Defects Zinc Fingers angiogenesis base cardiac angiogenesis chemical reaction cysteine sulfinic acid design endoplasmic reticulum stress genome-wide heart function in vivo inorganic phosphate meetings methionylmethionine mortality mouse development mouse model new therapeutic target overexpression oxidation protein function response ubiquitin ligase ubiquitin-protein ligase

项目摘要

DESCRIPTION (provided by applicant): The N-end rule pathway is one ubiquitin proteolytic pathway that relates the in vivo half-life of a protein to the identity of its N-terminal residue. Conjugation of arginine (Arg) from Arg-tRNAArg to N-terminal aspartate (Asp), glutamate (Glu), or cysteine (Cys) is part of this proteolytic pathway in that it can lead to ubiquitination of the resulting Arg-conjugated proteins. We have previously identified the mammalian Ate1 gene encoding Arg-transferases responsible for all known protein arginylation activities and have shown that Ate1-/- embryos die owing to various cardiovascular defects including ventricular hypoplasia, ventricular septal defect, and late angiogenesis. These results suggest that Ate1-dependent proteolysis of unknown substrate(s) is a crucial regulatory mechanism for myocardial growth and blood vessel integrity/maturation. However, the exact nature of the cardiovascular defects and the underlying molecular mechanisms remain elusive. Genomewide functional proteomic approach led us to identify a set of cardiovascular regulators (Rgs4, Rgs5, and Rgs16) as substrates of Ate1-dependent arginylation that may underlie, at least partially, Ate1-dependent cardiovascular homeostasis. Notably, Rgs4 and Rgs5 are GTPase-activating proteins (GAP) that act as negative regulators of GPCR-coupled Ga subunits and have been implicated as important regulators of Gq/Gi-activated signaling for myocardial growth and vascular maturation/integrity, respectively. Biochemical analyses showed that degradation of these substrates depends on the Cys2 residue as a degradation determinant, which is exposed to the N-terminus through cleavage of N-terminal Met by Met aminopeptidases. In the presence of sufficient oxygen (O2) and nitric oxide (NO), N-terminal Cys2 appears to be oxidized to CysO2 to create a structural homolog of Asp, an arginylation-permissive residue. The N-terminal Arg residue of arginylated RGS proteins is subsequently bound by specific E3 ligases whose identities remain unclear. Using an affinity-based proteomic approach, we isolated a set of E3 family (named Ubr1 through Ubr7) and demonstrated that Ubr1, Ubr2, and Ubr4 are the major E3s specific for protein arginylation and that Ubr1-/-Ubr2-/- and Ubr4-/- embryos die of cardiovascular defects. Based on these results, we hypothesize that the functions of Rgs4, Rgs5, and Rgs16 are modulated through the MetAPs-O2/NO-Ate1-Ubr proteolytic cascade. In Aim 1, we will characterize the physiological function of Ate1-dependent arginylation in cardiovascular development and signaling using tissue-specific Ate1 knockout mice in combination with transgenic mice overexpressing Gq in the heart. In Aim 2, to understand the molecular principles underlying Ate1-dependent cardiovascular development, we will characterize arginylation-dependent turnover and cotranslational modifications of Cys2 of Rgs4 and Rgs5. In Aim 3, as part of our long-term efforts to characterize ubiquitin ligases specific of arginylated substrates, we will characterize cardiovascular development of mice lacking Ubr4, a newly identified recognition component downstream of protein arginylation. 1

描述（由申请人提供）：N端规则途径是一种泛素蛋白水解途径，它将蛋白质的体内半衰期与其N端残基的身份相关联。 Arg-tRNAArg 中的精氨酸 (Arg) 与 N 末端天冬氨酸 (Asp)、谷氨酸 (Glu) 或半胱氨酸 (Cys) 的缀合是该蛋白水解途径的一部分，因为它可以导致所得 Arg 缀合蛋白泛素化。我们之前已经鉴定出哺乳动物 Ate1 基因编码负责所有已知蛋白质精氨酸化活性的精氨酸转移酶，并表明 Ate1-/- 胚胎由于各种心血管缺陷而死亡，包括心室发育不全、心室间隔缺损和晚期血管生成。这些结果表明，Ate1 依赖性未知底物的蛋白水解是心肌生长和血管完整性/成熟的关键调节机制。然而，心血管缺陷的确切性质和潜在的分子机制仍然难以捉摸。全基因组功能蛋白质组学方法使我们确定了一组心血管调节因子（Rgs4、Rgs5 和 Rgs16）作为 Ate1 依赖性精氨酸化的底物，可能至少部分是 Ate1 依赖性心血管稳态的基础。值得注意的是，Rgs4 和 Rgs5 是 GTP 酶激活蛋白 (GAP)，它们充当 GPCR 偶联 Ga 亚基的负调节因子，并分别作为心肌生长和血管成熟/完整性的 Gq/Gi 激活信号传导的重要调节因子。生化分析表明，这些底物的降解取决于作为降解决定因素的 Cys2 残基，该残基通过 Met 氨肽酶切割 N 端 Met 暴露到 N 端。在存在足够的氧气 (O2) 和一氧化氮 (NO) 的情况下，N 末端 Cys2 似乎会被氧化为 CysO2，从而产生 Asp 的结构同系物（一种允许精氨酸化的残基）。精酰化 RGS 蛋白的 N 端精氨酸残基随后被特定的 E3 连接酶结合，但其身份仍不清楚。使用基于亲和力的蛋白质组学方法，我们分离了一组 E3 家族（命名为 Ubr1 至 Ubr7），并证明 Ubr1、Ubr2 和 Ubr4 是对蛋白质精氨酸化具有特异性的主要 E3，并且 Ubr1-/-Ubr2-/- 和 Ubr4 -/- 胚胎死于心血管缺陷。基于这些结果，我们假设 Rgs4、Rgs5 和 Rgs16 的功能是通过 MetAPs-O2/NO-Ate1-Ubr 蛋白水解级联调节的。在目标 1 中，我们将使用组织特异性 Ate1 敲除小鼠与心脏中过表达 Gq 的转基因小鼠相结合，描述 Ate1 依赖性精氨酰化在心血管发育和信号传导中的生理功能。在目标 2 中，为了了解 Ate1 依赖性心血管发育的分子原理，我们将描述 Rgs4 和 Rgs5 的 Cys2 的精氨酰化依赖性周转和共翻译修饰。在目标 3 中，作为我们表征精氨酸化底物特异性泛素连接酶的长期努力的一部分，我们将表征缺乏 Ubr4 的小鼠的心血管发育，Ubr4 是一种新发现的蛋白质精氨酸化下游的识别成分。 1

项目成果

期刊论文数量（27）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Degradation of misfolded proteins in neurodegenerative diseases: therapeutic targets and strategies.

神经退行性疾病中错误折叠蛋白的降解：治疗目标和策略。

DOI：
发表时间：
2015-03-13
期刊：
影响因子：
12.8
作者：
Ciechanover, Aaron;Kwon, Yong Tae
通讯作者：
Kwon, Yong Tae

Pharmacological Modulation of the N-End Rule Pathway and Its Therapeutic Implications.

N 端规则途径的药理学调节及其治疗意义。

DOI：
发表时间：
2015-11
期刊：
影响因子：
13.8
作者：
Lee, Jung Hoon;Jiang, Yanxialei;Kwon, Yong Tae;Lee, Min Jae
通讯作者：
Lee, Min Jae

Role of the IL-6-JAK1-STAT3-Oct-4 pathway in the conversion of non-stem cancer cells into cancer stem-like cells.

IL-6-JAK1-STAT3-Oct-4 通路在非干癌细胞转化为癌症干细胞样细胞中的作用。

DOI：
10.1016/j.cellsig.2013.01.007
发表时间：
2013-04
期刊：
Cellular signalling
影响因子：
4.8
作者：
Kim SY;Kang JW;Song X;Kim BK;Yoo YD;Kwon YT;Lee YJ
通讯作者：
Lee YJ

UBR2 of the N-end rule pathway is required for chromosome stability via histone ubiquitylation in spermatocytes and somatic cells.

N 端规则途径的 UBR2 通过精母细胞和体细胞中的组蛋白泛素化来维持染色体稳定性。

DOI：
发表时间：
2012
期刊：
影响因子：
3.7
作者：
An, Jee Young;Kim, Euna;Zakrzewska, Adriana;Yoo, Young Dong;Jang, Jun Min;Han, Dong Hoon;Lee, Min Jae;Seo, Jai Wha;Lee, Yong Jun;Kim, Tae;de Rooij, Dirk G;Kim, Bo Yeon;Kwon, Yong Tae
通讯作者：
Kwon, Yong Tae

The N-end rule proteolytic system in autophagy.

自噬中的 N 端规则蛋白水解系统。

DOI：
发表时间：
2013-07
期刊：
影响因子：
13.3
作者：
Kim, Sung Tae;Tasaki, Takafumi;Zakrzewska, Adriana;Yoo, Young Dong;Sa Sung, Ki;Kim, Su;Cha;Hwang, Joonsung;Kim, Kyoung A;Kim, Bo Yeon;Kwon, Yong Tae
通讯作者：
Kwon, Yong Tae