ORB GENE REGULATION OF TRANSLATION

ORB基因翻译调控

• 批准号: 8171471
• 负责人: Paul D Schedl
• 金额: $ 0.24万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-09-01 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8171471
• 关键词: Axon; Binding; Biochemical; Candidate Disease Gene; Cell Nucleus; Cells; Complex; Computer Retrieval of Information on Scientific Projects Database; Coupled; Dendrites; Deposition; Development; Drosophila genus; Elements; Embryo; Ensure; Eukaryota; Event; Family; Feedback; Funding; Gene Expression Regulation; Genes; Genetic; Genetic Transcription; Genetic Translation; Goals; Grant; Growth; Homeostasis; Institution; Learning; Location; Masks; Mating Types; Memory; Messenger RNA; Microtubule-Associated Proteins; Movement; Mutation; Neurons; Nurses; Oocytes; Ovarian; Ovary; Play; Process; Protein Biosynthesis; Proteins; RNA-Binding Proteins; Research; Research Personnel; Resources; Role; Signal Transduction; Site; Somatic Cell; Source; Synaptic plasticity; Testing; Translating; Translational Activation; Translational Regulation; Translations; United States National Institutes of Health; Untranslated Regions; Yeasts; daughter cell; egg; fly; in vivo; member; mutant; prevent; segregation

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. mRNA localization plays a pivotal role in the establishment of polarity in the Drosophila egg and embryo. It typically begins with the transcription and packaging of the localized mRNAs in RNP complexes in nurse cell nuclei. These complexes 'mask' the mRNAs, preventing them from being translated while in transit, and 'mark' them for localization. The masked mRNAs are transported through the nurse cells and deposited in the oocyte. Depending upon their cis-acting localization elements, the mRNAs are then targeted to specific locations in the oocyte where they are anchored to the cytosketal network. Translational activation of the localized message provides a spatially restricted source of the protein product. Alternatively the mRNAs can be stored in a masked form until an appropriate signal, such as egg deposition, activates translation. This provides a mechanism for coordinating the localized synthesis of the protein with other developmental events. While the importance of mRNA localization (coupled with 'on site' translational regulation) was first documented in fly ovaries and early embryos, it is now clear that this regulatory mechanism is employed in many different contexts and occurs in virtually every eukaryote. For example, in yeast, the asymmetric segregation of ash-1 mRNA to the daughter cell provides a mechanism for ensuring that mating type switching does not occur in that cell. mRNA localization in somatic cells also occurs during the processes of movement, differentiation or growth. In neuronal cells, mRNAs encoding MAP2, a dendrite-specific microtubule-associated protein, and aCaMKII accumulate in the dendrites, but not in the axons. Localized mRNA translation has also been implicated in synaptic plasticity and learning and memory. In all of these cases, it is likely that the steps involved in mRNA localization and translational regulation are similar in broad outline to those described above in fly ovaries The proposed studies focus on the Drosophila orb gene which is one of the key components of the fly ovarian mRNA localization machinery. Orb is a founding member of the highly conserved CPEB family of RRM RNA binding proteins. In previous studies, we have shown that orb functions at the last step in the mRNA localization process, regulating the translation of the mRNA once it is on site. Orb binds to the 3? UTR of many localized mRNAs in vivo oskar (osk) Bicudal-D (Bic-D), and K(10) and is required to promote the translation of these mRNAs. In addition, Orb protein autoregulates its own expression by binding to the 3? UTR of localized orb mRNA and activating translation. This positive feedback loop is critical for the proper expression of Orb in the developing oocyte. The goal of our studies is to better understand how Orb autoregulates its own expression and how it controls the on site translation of mRNAs encoding factors critical for establishing oocyte/nurse cell identity and defining the polarity of the egg chamber and embryo. Part of the answer to these questions will come from identifying and characterizing other genes that are important for Orb autoregulation and/or function. For this purpose we propose to isolate proteins that are associated with Orb in vivo and identify them by Mass Spec analysis. To validate these biochemical studies we have devised a sensitized orb mutant background. The sensitized background will permit us to test mutations in candidate genes which encode proteins that are associated Orb in vivo for genetic interactions with Orb.

该副本是利用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这不一定是调查员的机构。 mRNA定位在果蝇卵和胚胎的极性中起关键作用。 它通常始于护士细胞核中RNP复合物中局部mRNA的转录和包装。这些复合物将mRNA“掩盖”，阻止它们在运输过程中翻译，并“标记”它们进行本地化。 掩盖的mRNA通过护士细胞运输并沉积在卵母细胞中。根据其顺式作用的定位元素，mRNA随后被靶向卵母细胞中的特定位置，将它们固定在Cytosketal网络上。 局部消息的翻译激活提供了蛋白质产物的空间限制来源。 或者，可以将mRNA以掩盖的形式存储，直到适当的信号（例如鸡蛋沉积）激活翻译。这提供了一种协调蛋白质与其他发育事件的局部合成的机制。 虽然首先在卵巢和早期胚胎中记录了mRNA定位的重要性（与“现场”翻译法规结合在一起），但现在很明显，这种调节机制在许多不同的情况下都采用，并且实际上是在所有真核生物中发生的。例如，在酵母中，Ash-1 mRNA与子细胞的不对称分离提供了确保该细胞中未发生交配类型切换的机制。 在运动，分化或生长过程中，在体细胞中的MRNA定位也发生。 在神经元细胞中，编码MAP2的mRNA，树突特异性微管相关蛋白和AcamkII积聚在树突中，但在轴突中却没有。 局部mRNA翻译也与突触可塑性，学习和记忆有关。 在所有这些情况下，mRNA定位和翻译调节中涉及的步骤在广泛的轮廓上可能与上述苍蝇卵巢中描述的步骤相似 拟议的研究集中于果蝇球基因，这是卵巢mRNA定位机制的关键组成部分之一。 ORB是RRM RNA结合蛋白的高度保守CPEB家族的创始成员。 在先前的研究中，我们已经表明，ORB在mRNA定位过程的最后一步中的功能，一旦在现场，它调节了mRNA的翻译。球与3结合？许多局部mRNA在体内OSKAR（OSK）BICUDAL-D（BIC-D）和K（10）的UTR，并且需要促进这些mRNA的翻译。 另外，Orb蛋白通过与3结合来自动调节其自身的表达？局部球mRNA的UTR和激活翻译。这种积极的反馈回路对于在发育中的卵母细胞中正确表达球体至关重要。 我们的研究的目的是更好地了解Orb如何自动调节其自身的表达以及它如何控制MRNA编码因子的现场翻译，这对于建立卵母细胞/护士细胞身份至关重要，并定义了鸡蛋室和胚胎的极性。这些问题的答案的一部分将来自识别和表征其他对球体自动调节和/或功能很重要的基因。 为此，我们建议分离与体内球体相关的蛋白质，并通过质量规格分析识别它们。 为了验证这些生化研究，我们设计了一种敏化的球突变体背景。 敏化背景将使我们能够在候选基因中测试突变，这些突变编码与体内相关的球体与球体相互作用的蛋白质。