Regulated RNA degradation is required for Drosophila oogenesis

果蝇卵子发生需要调节 RNA 降解

• 批准号: 9470415
• 负责人: Patrick Blatt
• 金额: $ 2.87万
• 依托单位: STATE UNIVERSITY OF NEW YORK AT ALBANY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-05 至 2019-09-04
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9470415
• 关键词: Address; Adult; Binding Proteins; Biochemistry; Bioinformatics; Code; Complex; Confocal Microscopy; Coupled; Data; Defect; Degradation Pathway; Deposition; Detection; Development; Developmental Gene; Disease; Double-Stranded RNA; Drosophila genus; Drosophila melanogaster; Elements; Ensure; Exhibits; Female; Female sterility; Fertility; Gene Targeting; Generations; Genes; Genetic; Germ Cells; Germ Lines; Goals; Gonadal structure; Homologous Gene; Human; Immunohistochemistry; Infertility; Lead; Length; Link; Mediating; Messenger RNA; Metabolism; Modeling; Mothers; Non-Stop Decay; Nonsense-Mediated Decay; Oocytes; Oogenesis; Open Reading Frames; Organism; Ovary; Parents; Pathway interactions; Pattern; Phenocopy; Phenotype; Play; Process; Production; Proteins; Quality Control; RNA; RNA Degradation; RNA Helicase; RNA-Binding Proteins; Recruitment Activity; Regulation; Reporter; Research; Role; SKIV2L gene; Small RNA; Somatic Cell; Specific qualifier value; Spermatogenesis; Syndrome; Testing; Tissue-Specific Gene Expression; Transcript; Up-Regulation; Work; Yeasts; differential expression; egg; exosome; fly; genetic information; insight; mRNA Decay; mRNA Surveillance; mRNA Transcript Degradation; male; member; mutant; neuronal cell body; next generation; offspring; overexpression; prevent; protein complex; reduce symptoms; scaffold; therapy design; transcriptome; transcriptome sequencing

Abstract The germ line generates gametes that link generations by passing genetic information from parent to offspring. During oogenesis, the egg receives critical mRNAs from the mother that help launch the next generation; any mistakes in this process could be detrimental to the offspring. However, it is not known if the maternally provided mRNAs are surveilled for quality or how aberrantly expressed mRNAs impact oogenesis. Twister (Tst), an RNA helicase conserved from yeast to humans, promotes decay of defective mRNAs. Loss of the tst human homolog, SKIV2L, results in trichohepatoenteric (THE) syndrome, which has no known treatment. Tst is part of the Ski complex that unwinds RNA duplexes, which are then subject to degradation by the exosome complex. mRNA targets of quality control mechanisms such as nonsense-mediated decay (NMD), no-go decay (NGD) and nonstop decay (NSD) are substrates for the Ski complex and the exosome. We find that the tst homozygous flies are viable but female sterile, exhibiting an oogenesis defect. We find tst is required in the germ line but not the soma of the gonad for proper oogenesis. Using RNA-Seq and bioinformatic analyses, we found that in tst mutants most of the transcriptome is not affected, but a distinct subset is upregulated and only a few transcripts are down regulated. This is consistent with Tst playing a central role in RNA degradation. The transcripts upregulated in tst mutants are poorly expressed in wild type conditions and these mRNAs are typically shorter in their 5’UTR, ORF and 3’UTR regions. Surprisingly, we find that the Tst targets do not have features of the canonical mRNA decay pathways such as NMD, NGD and NSD. We hypothesize that Tst plays a central role in surveilling the mRNAs supplied to the developing oocyte and facilitates the turnover of developmentally expressed mRNAs through their 3’UTRs. The main goals of this research are to: 1) determine how mRNA degradation mediated by Tst promotes proper oogenesis, 2) identify mRNAs targets of Tst, and 3) elucidate the mechanism by which Tst is recruited to these mRNA targets to initiate their degradation. These studies aim to establish the contributions of Tst to egg development and female fertility through turnover of developmentally regulated mRNAs. If Tst mediates turnover of developmentally regulated transcripts, this can provide insight to diseases such as THE. Our work in a genetically tractable organism will allow us to elucidate a target to design treatments against to alleviate symptoms resulting from insufficient mRNA metabolism and turnover. !

抽象的 种系会产生游戏，通过将遗传信息从父母传递到 后代。在卵子发生过程中，鸡蛋从母亲那里收到关键的mRNA，有助于推出下一个 一代;此过程中的任何错误都可能对后代有害。但是，尚不知道是否 对母体提供的mRNA的质量或异常表达的mRNA撞击卵子发生的方式进行了调查。 Twister（TST）是一种从酵母到人类的RNA解旋酶，促进了有缺陷的mRNA衰减。损失 TST人类同源物SKIV2L导致Trichohepatoenteric（The）综合征，尚无已知 TST是放松RNA双链体的滑雪综合体的一部分，然后会降解 外泌体复合物。质量控制机制的mRNA靶标，例如废话介导的衰减 （NMD），No-Go Decay（NGD）和NONSTOP衰减（NSD）是滑雪络合物和外部体的底物。 我们发现，TST纯合蝇是可行的，但雌性无菌，表现出卵子缺陷。我们找到了TST 使用RNA-seq和 生物信息学分析，我们发现在TST突变体中，大多数转录组不受影响，而是独特的 子集已更新，只有少数成绩单受到调节。这与TST播放是一致的 RNA降解中的中心作用。 TST突变体中更新的成绩单在野生型中表达不佳 条件和这些mRNA通常在其5'UTR，ORF和3'UTR地区较短。令人惊讶的是，我们发现 TST目标没有规范mRNA衰变途径的特征，例如NMD，NGD和 NSD。我们假设TST在监视提供给开发的mRNA方面起着核心作用 卵母细胞并促进了通过其3'utrs发达的mRNA的营业额。 这项研究的主要目标是：1）确定TST介导的mRNA降解如何促进适当 2）识别TST的mRNA靶标，3）阐明了TST募集到这些靶标的机制 mRNA靶标发起其降解。这些研究旨在建立TST对鸡蛋的贡献 发育和女性生育能力通过开发的受监管的mRNA的营业率。如果TST进行调解 发展为受管制的成绩单的营业额，这可以为诸如疾病等疾病提供见解。我们的工作 在一般可牵引的有机体中，我们可以阐明目标，以设计治疗方法 mRNA代谢和营业额不足导致的症状。 呢