Examination of Ornithine Decarboxylase Antizyme RNA Structure and Function from Various Organisms for the Development of Antibiological Agents

检查不同生物体的鸟氨酸脱羧酶抗酶 RNA 结构和功能，用于开发抗生素

• 批准号: 10730595
• 负责人: JULIANE K STRAUSS-SOUKUP
• 金额: $ 44.1万
• 依托单位: CREIGHTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-01 至 2026-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10730595
• 关键词: Abbreviations; Acylation; Affect; Affinity; Anabolism; Animals; Antibiotics; Antifungal Agents; Antineoplastic Agents; Bacteria; Binding; Biological Assay; Bypass; Calorimetry; Cell Culture System; Cell Differentiation process; Cell Proliferation; Cells; Characteristics; Complex; DNA; Development; Dialysis procedure; Disease; Drug Targeting; Elements; Enzymes; Equilibrium; Feedback; Gene Expression; Gene Expression Regulation; Genes; Genetic Transcription; Germ Cells; Homeostasis; Human; Hydroxyl Radical; In Vitro; Insecta; Invertebrates; Ligand Binding; Ligands; Luciferases; Malignant Neoplasms; Malignant neoplasm of prostate; Mammalian Cell; Mammals; Measures; Mentorship; Messenger RNA; Metabolic; Metabolism; Mus; Names; Nucleotides; Organism; Ornithine Decarboxylase; Pathway interactions; Pesticides; Phospholipids; Plants; Play; Polyamines; Primer Extension; Process; Production; Proteins; Publications; Putrescine; RNA; RNA Sequences; Reporter Genes; Ribosomal Frameshifting; Roentgen Rays; Role; Signal Transduction; Site; Skin Cancer; Specificity; Spermidine; Spermine; Structure; Students; Supporting Cell; Testis; Titrations; Translations; Untranslated RNA; Work; X-Ray Crystallography; analog; anti-cancer; cell growth; design; drug development; flexibility; fungus; gene product; malignant breast neoplasm; new therapeutic target; novel; ornithine decarboxylase antizyme; overexpression; pathogenic fungus; premature; sensor; three dimensional structure; three-dimensional modeling; tumor; undergraduate research; uptake

PROJECT SUMMARY Nearly all organisms possess the capability to synthesize the natural polyamines - putrescine, spermidine and spermine - which are essential for cell growth and differentiation. Due to the ability of polyamines to interact with nearly every biomolecule - DNA, RNA, phospholipids, proteins and ATP, to name a few - they play many roles within the cell in order to support cell growth. It has been well documented that polyamine levels in mammalian cells correlate with the rate of cell growth, high polyamine concentrations have been observed in rapidly proliferating cells and low concentrations have been measured in slow-growing or quiescent cells. Not surprisingly, the transport and metabolism of polyamines are highly regulated by complex feedback mechanisms. Ornithine decarboxylase (ODC) is the key regulatory enzyme in polyamine biosynthesis. ODC homeostasis affects cell growth and cancer development. ODC over-expression has been observed in many tumor types, including prostate, breast, and skin cancers. Both ODC and cellular uptake of polyamines is inhibited by Ornithine Decarboxylase Antizyme (OAZ). The making of Antizyme protein from OAZ mRNA requires translational frameshifting at a highly conserved site to bypass premature termination. Mammalian OAZ mRNAs further possess a pseudoknot (PK) RNA 3¢ to the frameshift site that stimulates +1 frameshifting. Moreover, frameshifting is stimulated by polyamines, thus providing a feedback mechanism whereby the accumulation of metabolic products inhibits biosynthesis. Although the role of the OAZ pseudoknot RNA element (further designated OAZ-PK) in polyamine-dependent frameshifting has been investigated, it has not been examined as a distinct polyamine “sensor”. Riboswitches are elements within noncoding regions of mRNAs that directly bind to cellular metabolites and modulate gene expression. Many riboswitches provide a mechanism of feedback regulation for gene products within the biosynthetic pathway of the cognate metabolite. Riboswitches are widespread among bacteria, and one class further resides in fungi and plants, but no riboswitches have been found in animals. It is proposed that the OAZ-PK RNA functions as a riboswitch, and herein evidence is provided that this noncoding RNA is a polyamine sensor. This RNA element is highly conserved among vertebrate genes required for spermine biosynthesis. Development of drugs that target putative spermine riboswitches from different organisms might therefore be used for wide-ranging purposes such as anticancer agents, antifungal agents, or pesticides. This proposal will examine the structure and function of the OAZ-PK RNA from various organisms, including those of biomedical relevance – human, pathogenic fungi and disease harboring insects - with the following specific aims: (1) examine the specificity and affinity of polyamine binding to OAZ RNAs from various organisms, (2) investigate the three-dimensional structure of OAZ RNAs, and (3) explore the role of Antizyme OAZ RNAs from various organisms in control of gene expression.

项目概要 几乎所有生物体都具有合成天然多胺的能力 - 腐胺、亚精胺和 精胺 - 由于多胺具有相互作用的能力，对细胞生长和分化至关重要。 几乎每一种生物分子——DNA、RNA、磷脂、蛋白质和 ATP 等等——它们都发挥着多种作用 有充分证据表明，多胺水平在细胞内发挥支持细胞生长的作用。 哺乳动物细胞与细胞生长速率相关，已观察到高多胺浓度 在生长缓慢或静止的细胞中测量到快速增殖的细胞和低浓度。 令人惊讶的是，多胺的运输和代谢受到复杂反馈的高度调节 鸟氨酸脱羧酶（ODC）是多胺生物合成的关键调节酶。 在许多细胞中观察到稳态会影响细胞生长和癌症发展。 肿瘤类型，包括前列腺癌、乳腺癌和皮肤癌，ODC 和多胺的细胞摄取均受到影响。 被鸟氨酸脱羧酶抗酶 (OAZ) 抑制 从 OAZ mRNA 制备抗酶蛋白。 需要在高度保守的位点进行平移移码以避免哺乳动物的过早终止。 OAZ mRNA 还具有位于移码位点的假结 (PK) RNA 3 分，可刺激 +1 移码。 此外，移码是由多胺刺激的，从而提供了反馈机制，使 代谢产物的积累抑制了OAZ假结RNA的生物合成。 多胺依赖性移码中的元件（进一步指定为 OAZ-PK）已被研究，但尚未 核糖开关被认为是一种独特的多胺“传感器”，是非编码区域内的元件。 直接与细胞代谢物结合并调节基因表达的 mRNA 许多核糖开关提供了一个 同源生物合成途径中基因产物的反馈调节机制 核糖开关在细菌中广泛存在，一类还存在于真菌和植物中，但是 在动物体内未发现核糖开关，有人提出 OAZ-PK RNA 具有核糖开关的功能。 并且本文提供的证据表明该非编码RNA是多胺传感器。 精胺生物合成所需的脊椎动物基因中的保守性 开发靶向药物。 因此，来自不同生物体的假定精胺核糖开关可能具有广泛的用途 例如抗癌剂、抗真菌剂或杀虫剂。该提案将检查其结构和特性。 来自各种生物体的 OAZ-PK RNA 的功能，包括具有生物医学相关性的生物体 – 人类、 含有昆虫的病原真菌和疾病 - 具有以下具体目标：（1）检查特异性 以及多胺与来自不同生物体的 OAZ RNA 结合的亲和力，(2) 研究三维 OAZ RNA 的结构，以及 (3) 探索来自不同生物体的抗酶 OAZ RNA 在控制 基因表达。