Translation regulation of gene expression in toxic dinoflagellates

有毒甲藻基因表达的翻译调控

• 批准号: 8550056
• 负责人: ROSEMARY JAGUS
• 金额: $ 13.14万
• 依托单位: CENTER FOR ENVIRONMENTAL SCIENCE
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-24 至 2017-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8550056
• 关键词: Affect; Affinity; Affinity Chromatography; Antibodies; Binding; Biological Assay; Biological Markers; Cell Extracts; Cells; Cellular Stress; Complementary DNA; Coupled; Databases; Development; Dinophyceae; Environment; Eukaryota; Family member; Gel; Gene Expression; Gene Expression Regulation; Genes; Genetic; Genome; Genomics; Goals; Growth; In Vitro; Incubated; Intervention; Investigation; Length; Life; Life Cycle Stages; Light; Messenger RNA; Monitor; Mus; Neurotoxins; Organism; Parasites; Peroxides; Phase; Phosphorylation; Phosphotransferases; Physiological; Plants; Plasma; Plasmodium falciparum; Play; Policies; Polyadenylation; Process; Protein Biosynthesis; Proteins; Protista; Recombinant Proteins; Recombinants; Regulation; Research Personnel; Role; Site; Spliced Leader Sequences; Structure; Surface; Toxic effect; Toxin; Toxoplasma gondii; Trans-Splicing; Transcriptional Regulation; Translational Regulation; Translations; Variant; Yeasts; cDNA Library; eIF2 Regulation Pathway; harmful algal blooms; in vitro activity; in vivo; innovation; inorganic phosphate; insight; member; public health relevance; red tide; response; screening; sound; stressor

DESCRIPTION (provided by applicant) A number of dinoflagellate species are known to produce potent neurotoxins. Their blooms are commonly referred to as ''red tides.'' An understanding of the genetic regulatory mechanisms that affect bloom growth and, in particular, the development of biomarkers to assess bloom growth status is essential for the development of scientifically sound management and mitigation policies. The genetic organization and regulation of these organisms has been shown to be distinct from non-protist eukaryotes. Dinoflagellates are remarkable for their extremely large genomes that show little transcriptional regulation and with genes present as multiple tandem copies that are polycistronically processed through coupled trans-splicing and polyadenylation. A broad range of investigations has implicated mRNA recruitment as a major site of regulation of gene expression. However, relatively little is understood regarding translational initiation and its regulation in these organisms. The goal of this application is to unravel the roles of the likely key players in translational regulation, eIF2 and eIF4, along with the spliced leader cap structures in regulating gene expression. Using the icthyotoxic dinoflagellate, Karlodinium veneficum and the toxin producing Karenia brevis, the investigators will a) determine whether changes in eIF2-alpha phosphorylation underlie dial changes in gene expression or responses to different stressors; b) begin characterization of the eIF2-alpha-kinases from K. veneficum; c) characterize the ability of K.veneficum eIF4E family member to bind to cap structures and translational binding partners; d) assess the role of K.veneficum eIF4E members in mRNA recruitment. The investigators current understanding of translational regulation is derived mainly from studies in mouse, yeast and plants and does not allow for the diversity of eukaryotic life, the bulk of which is to be found in the Protista. The investigators' results are expected to define a new paradigm for translational regulation in dinoflagellates and represent an innovative approach to increase our understanding of the translational process itself as well as to bridge the gap between genomics and physiological complexity in dinoflagellates. Given the central role of translation in dinoflagellate gene expression, this will provide critical insight into mechanisms relevant to dinoflagellate growth, toxicity, and the regulation of harmful algal blooms. Public Health Relevance: A number of dinoflagellate species are known to produce potent neurotoxins. Their blooms are commonly referred to as ''red tides.'' The investigators are studying the mechanisms by which these organisms can regulate their growth and respond to the environment. Because of their unique genome structure, dinoflagellates regulate themselves in unusual ways. The studies proposed here will provide critical insight into mechanisms relevant to dinoflagellate growth, toxicity, and the regulation of harmful algal blooms, with a lon term goal of providing avenues for intervention.

描述（由申请人提供） 已知许多甲藻物种产生强效神经毒素。它们的爆发通常被称为“赤潮。了解影响水华生长的遗传调控机制，特别是开发生物标志物来评估水华生长状况，对于制定科学合理的管理和缓解政策至关重要。这些生物的遗传组织和调节已被证明是不同于非原生真核生物。 甲藻是显着的，其非常大的基因组，显示很少的转录调控，并与基因作为多个串联拷贝，通过耦合的反式剪接和多聚腺苷酸化多顺反子加工。广泛的研究表明，mRNA募集是基因表达调控的主要位点。然而，对这些生物体中的翻译起始及其调控了解相对较少。本申请的目的是阐明翻译调控中可能的关键参与者eIF 2和eIF 4的作用，沿着剪接的前导帽结构调节基因表达。研究人员将利用具有鱼刺毒性的甲藻Karlodinium veneficum和产生毒素的Karenia brevis，a）确定eIF 2-α磷酸化的变化是否是基因表达或对不同应激源反应的变化的基础; B）开始表征来自K. veneficum; c）表征K.veneficum eIF 4 E家族成员结合帽结构和翻译结合配偶体的能力;研究人员目前对翻译调控的理解主要来自小鼠，酵母和植物的研究，不考虑真核生物的多样性，其中大部分在原生生物中发现。研究人员的研究结果有望为甲藻的翻译调控定义一个新的范式，并代表了一种创新的方法，以增加我们对翻译过程本身的理解，并弥合基因组学和生理复杂性之间的差距。考虑到翻译在甲藻基因表达中的核心作用，这将 对甲藻生长、毒性和有害藻华的调节机制提供重要的见解。 公共卫生相关性：已知一些甲藻物种产生强效神经毒素。它们的爆发通常被称为“赤潮。研究人员正在研究这些生物体调节其生长和对环境作出反应的机制。由于其独特的基因组结构，甲藻以不寻常的方式调节自己。这里提出的研究将提供关键的洞察有关甲藻生长，毒性和有害藻华的调节机制，提供干预途径的长期目标。