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.

项目摘要 几乎所有生物都具有合成天然多胺的能力 - 腐烂，精子和 精子 - 对于细胞生长和分化至关重要。由于多胺相互作用的能力 几乎每一个生物分子 - DNA，RNA，磷脂，蛋白质和ATP，仅举几例 细胞内的作用以支持细胞生长。已经有充分的文献记录了多胺水平 哺乳动物细胞与细胞生长速率相关，在 快速增殖的细胞和低浓度已在缓慢生长或静止的细胞中测量。不是 令人惊讶的是，多胺的运输和代谢受复杂反馈的高度调节 机制。鸟氨酸脱羧酶（ODC）是多胺生物合成中的关键调节酶。 ODC 稳态会影响细胞生长和癌症的发展。在许多人中都观察到了ODC过表达 肿瘤类型，包括前列腺，乳房和皮肤癌。多胺的ODC和细胞摄取均为 鸟氨酸脱羧酶抗酶（OAZ）抑制。从OAZ mRNA制造抗酶蛋白 需要在高度保守的地点进行翻译框架以绕过过早终止。哺乳动物 OAZ mRNA进一步具有伪码位刺激+1帧速率的伪毫移位位置3¢。 此外，多胺刺激了帧速率，从而提供了一种反馈机制 代谢产物的积累抑制了生物合成。虽然OAZ Pseudoknot RNA的作用 已经研究了依赖多胺依赖性帧的元素（进一步指定的OAZ-PK），已经研究 被检查为独特的多胺“传感器”。核糖开关是非编码区域内的元素 直接结合细胞代谢物并调节基因表达的mRNA。许多核糖开关提供了 同源的生物合成途径内基因产物的反馈调节机制 代谢物。核糖开关在细菌中普遍存在，一类进一步居住在真菌和植物中，但 在动物中没有发现核糖开关。建议OAZ-PK RNA充当核糖开关， 并在此证明该非编码RNA是多胺传感器。这个RNA元素高度 在精子生物合成所需的脊椎动物基因中保守。靶向药物的开发 因此，来自不同生物体的假定的精子核糖开关可能用于大范围 例如抗癌剂，抗真菌剂或农药。该建议将检查结构和 来自各种生物体的OAZ-PK RNA的功能，包括生物医学相关性的功能 - 人类， 病原真菌和具有昆虫的疾病 - 具有以下特定目的：（1）检查特异性 多胺与来自各种生物体的OAZ RNA结合的亲和力，（2）研究三维 OAZ RNA的结构和（3）探索来自各种生物体中的抗卵OAZ RNA的作用 基因表达。