Novel Cap Analogs and Interactions with Target Proteins

新型帽类似物以及与靶蛋白的相互作用

• 批准号: 6897495
• 负责人: ROBERT E. RHOADS
• 金额: $ 3.97万
• 依托单位: LOUISIANA STATE UNIV HSC SHREVEPORT
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-07-01 至 2006-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6897495
• 关键词: Caenorhabditis elegans; binding sites; chemical binding; messenger RNA; nuclear magnetic resonance spectroscopy; phosphorylation; posttranscriptional RNA processing; protein biosynthesis; protein isoforms; protein protein interaction; pyrophosphatase; ribosomal proteins; thermodynamics; tissue /cell culture; translation factor

DESCRIPTION (provided by applicant) This application for a Fogarty International Research Collaboration Award (FIRCA) is an extension of Grant R01GM20818, entitled "Regulation of Eukaryotic Protein Synthesis Initiation." The research will be done primarily in Poland at Warsaw University in collaboration with Edward Darzynkiewicz. The long-term goals of both the Parent Grant and the FIRCA project are to understand the steps that occur during the recruitment of eukaryotic mRNA to the ribosome and how they are regulated. This process determines both the overall rate of protein synthesis and also the spectrum of mRNAs translated. The four Specific Aims of the FIRCA application involve the 7-methylguanosine-containing cap of mRNA and the target protein which it interacts during cap-dependent initiation of protein synthesis, elF4E. The nematode C. elegans provides unparalleled advantages for studying such a complex biochemical process as initiation of protein synthesis. Surprisingly, it expresses five eIF4E isoforms, termed IFE proteins, that have differing cap-binding properties. In Specific Aim 1, we will synthesize and test in biological systems new classes of di- and oligonucleotide cap analogues, specifically a new series of anti-reverse cap analogues, dinucleotide cap analogues intended to be resistant to degradation by pyrophosphatases, and capped trinucleotides. In Specific Aim 2, we will synthesize and test new inhibitors of translation based on cap analogues that are able to cross the plasma membrane, in an attempt to target cap-dependent translation as an anticancer strategy. In Specific Aim 3, we will analyze thermodynamiC and solvent aspects of cap binding to elF4E isoforms of C. elegans. In Specific Aim 4, we will investigate the role of elF4E phosphorylation using chemically synthesized human Ser-209 phospho-elF4E. Because overexpression of elF4E causes malignant transformation of cells, and because naturally occurring tumors contain greatly elevated levels of elF4E, these studies have relevance to cancer, especially Specific Aim 2. They also have relevance for diseases caused by picornaviruses such as poliovirus (polio), rhinovirus (the common cold) and Coxsackievirus (heart failure), since picomaviruses shut down cap-dependent translation by proteolytically cleaving elF4G, the protein that tethers eIF4E to the 40S ribosomal subunit during initiation of translation.

描述（由申请人提供） 这项福格蒂国际研究合作奖 (FIRCA) 申请是 R01GM20818 拨款的延伸，题为“真核蛋白质合成起始的调节”。该研究将主要在波兰华沙大学与爱德华·达尔津凯维奇 (Edward Darzynkiewicz) 合作进行。 Parent Grant 和 FIRCA 项目的长期目标是了解真核 mRNA 招募到核糖体过程中发生的步骤以及它们是如何受到调节的。这个过程决定了蛋白质合成的总体速率以及翻译的 mRNA 的范围。 FIRCA 应用的四个具体目标涉及 mRNA 的含 7-甲基鸟苷帽及其在蛋白质合成的帽依赖性启动过程中相互作用的靶蛋白 eF4E。线虫线虫为研究蛋白质合成起始等复杂的生化过程提供了无与伦比的优势。令人惊讶的是，它表达五种 eIF4E 亚型，称为 IFE 蛋白，具有不同的帽结合特性。在具体目标 1 中，我们将在生物系统中合成并测试新类别的二核苷酸和寡核苷酸帽类似物，特别是一系列新的抗反向帽类似物、旨在抵抗焦磷酸酶降解的二核苷酸帽类似物以及加帽三核苷酸。在具体目标 2 中，我们将合成并测试基于能够穿过质膜的帽子类似物的新翻译抑制剂，试图将帽子依赖性翻译作为一种抗癌策略。在具体目标 3 中，我们将分析帽与线虫 eF4E 同种型结合的热力学和溶剂方面。在具体目标 4 中，我们将使用化学合成的人 Ser-209 磷酸化 eIF4E 研究 eIF4E 磷酸化的作用。因为 elF4E 的过度表达会导致细胞恶性转化，并且因为自然发生的肿瘤含有大大升高的 elF4E 水平，所以这些研究与癌症相关，尤其是特定目标 2。它们还与小核糖核酸病毒引起的疾病相关，例如脊髓灰质炎病毒（脊髓灰质炎）、鼻病毒（普通感冒）和柯萨奇病毒（心力衰竭），因为小核糖核酸病毒通过以下方式关闭帽依赖性翻译： 蛋白水解裂解 eIF4G，该蛋白在翻译起始过程中将 eIF4E 与 40S 核糖体亚基连接。

项目成果

New affinity resin for purification of cap-binding proteins.

用于纯化帽结合蛋白的新型亲和树脂。

• DOI: 10.1081/ncn-200061782
• 发表时间: 2005
• 期刊: Nucleosides, nucleotides & nucleic acids
• 作者: Jankowska-Anyszka,Marzena;Nogalski,Maciej;Darzynkiewicz,Edward
• 通讯作者: Darzynkiewicz,Edward

Thermodynamics of mRNA 5' cap binding by eukaryotic translation initiation factor eIF4E.

真核翻译起始因子 eIF4E 结合 mRNA 5 帽的热力学。

• DOI: 10.1021/bi0491651
• 发表时间: 2004
• 期刊: Biochemistry.
• 作者: Niedzwiecka,Anna;Darzynkiewicz,Edward;Stolarski,Ryszard
• 通讯作者: Stolarski,Ryszard

Novel dinucleoside 5',5'-triphosphate cap analogues. Synthesis and affinity for murine translation factor eIF4E.

新型二核苷 5,5-三磷酸帽类似物。

• DOI: 10.1081/ncn-200060103
• 发表时间: 2005
• 期刊: Nucleosides, nucleotides & nucleic acids.
• 作者: Stepinski,Janusz;Zuberek,Joanna;Jemielity,Jacek;Kalek,Marcin;Stolarski,Ryszard;Darzynkiewicz,Edward
• 通讯作者: Darzynkiewicz,Edward