Translation regulation of gene expression in toxic dinoflagellates

有毒甲藻基因表达的翻译调控

基本信息

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
项目类别：
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

项目摘要

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）已知许多鞭毛藻种类产生有效的神经毒素。它们的盛开通常被称为“红潮”。对影响开花生长的遗传调节机制的理解，尤其是评估Bloom生长状况的生物标志物的发展对于发展科学声音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与非统治者真核生物不同。 Winoflagellates对于其极大的基因组而言是显着的，这些基因组几乎没有转录调节，并且基因作为多个串联拷贝，这些副本是通过耦合的反式变速器和多腺苷酸化来进行多余的。广泛的研究已将mRNA募集成为基因表达调节的主要部位。但是，关于翻译起始及其在这些生物中的调节，相对较少的理解。该应用的目的是揭示可能主要参与者在翻译调节，EIF2和EIF4中的作用，以及剪接的领导者盖结构在调节基因表达中的作用。使用诊断性鞭毛鞭毛盐，karlodinium veneficum和产生karenia brevis的毒素，研究人员将a）确定基因表达的EIF2-Alpha磷酸化的变化是基因表达或对不同压力因子的反应的变化； b）开始表征K. veneficum的EIF2-α-激酶； c）表征K. veneficum eif4e家族成员与帽结构和翻译结合伙伴结合的能力； d）评估K. veneficum eIF4E成员在mRNA募集中的作用。研究者当前对翻译调节的理解主要来自小鼠，酵母和植物的研究，并且不允许真核生活的多样性，其中大部分都可以在蛋白质中找到。预计研究者的结果将定义一种新的鞭毛鞭毛调节范式，并代表了一种创新的方法，以增加我们对转化过程本身的理解，并弥合鞭毛鞭毛藻中基因组学和生理复杂性之间的差距。鉴于翻译在鞭毛藻基因表达中的核心作用，这将对与鞭毛藻生长，毒性和有害藻华的调节有关的机制提供了批判性的见解。公共卫生相关性：已知许多鞭毛藻物种会产生有效的神经毒素。他们的花朵通常被称为“红潮”。研究人员正在研究这些生物可以调节其生长并对环境反应的机制。由于其独特的基因组结构，Winoflagellates以异常的方式调节了自己。此处提出的研究将对与鞭毛鞭毛生长，毒性和有害藻类开花的调节有关的机制进行批判性洞察力，并具有提供干预途径的LON术语。