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） Fogarty国际研究合作奖（FIRCA）的申请是赠款R01GM20818的延伸，名为“真核蛋白质合成起始”。这项研究将主要在华沙大学与爱德华·达齐维奇（Edward Darzynkiewicz）合作。父母赠款和FIRCA项目的长期目标是了解在真核mRNA招募到核糖体以及如何调节的过程中发生的步骤。该过程决定了蛋白质合成的总体速率，也决定了被翻译的mRNA的光谱。 FIRCA施用的四个特定目的涉及mRNA的7-甲基鸟苷帽和在蛋白质合成蛋白质合成期间相互作用的靶蛋白ELF4E。线虫C.秀丽隐杆线虫为研究像蛋白质合成的开始这样的复杂生化过程提供了无与伦比的优势。令人惊讶的是，它表达了五个EIF4E同工型，称为IFE蛋白，具有不同的盖结合特性。在特定目标1中，我们将在生物系统中合成和测试新的DI-和寡核苷酸帽类似物，特别是一系列新的抗反向帽类似物，二核苷酸帽类似物，旨在耐pyRophophaspass和Trinucleotides抗性。在特定的目标2中，我们将基于能够越过质膜的帽类似物合成和测试新的翻译抑制剂，以试图将依赖性帽依赖性翻译作为抗癌策略。在特定的目标3中，我们将分析帽结合与秀丽隐杆线虫的ELF4E同工型的热力学和溶剂方面。在特定的目标4中，我们将使用化学合成的人类Ser-209磷酸化磷酸4e研究ELF4E磷酸化的作用。 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蛋白水解裂解ELF4G，这是在翻译开始期间将EIF4E推至40S核糖体亚基的蛋白质。

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