Novel functions for Sm-class RNAs in the regulation of gene expression

Sm 类 RNA 在基因表达调控中的新功能

• 批准号: 10330827
• 负责人: Demian Cazalla
• 金额: $ 38.38万
• 依托单位: UNIVERSITY OF UTAH
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-02-16 至 2027-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10330827
• 关键词: 3' Untranslated Regions; Base Pairing; Binding; Binding Sites; Biochemical; Biological Models; Cells; Code; Disease; Eukaryota; Gene Expression; Gene Expression Regulation; Genetic; Goals; Herpesviridae; Homeostasis; Laboratories; Link; Mammalian Cell; Mechanics; Mediating; Messenger RNA; MicroRNAs; Molecular Biology; Primates; RNA; RNA Binding; RNA Splicing; Research; Ribonucleoproteins; Saimiriine Herpesvirus 2; Seeds; Small Nuclear RNA; Spider Monkey; Techniques; Untranslated RNA; Untranslated Regions; Viral; Viral Cancer; Virus Diseases; Work; flexibility; gammaherpesvirus; genome-wide; in vivo; interest; mRNA Precursor; novel; particle; snRNP Structural Core Protein

Summary The overarching goal of my laboratory is to define the functions and mechanisms of a group of non- coding RNAs (ncRNAs): the small nuclear, U-rich RNAs (snRNAs). Cellular snRNAs associate with Sm proteins to form ribonucleoprotein particles (RNPs) that are essential components of cellular machineries responsible for the precursor messenger RNA (pre-mRNA) processing steps of splicing and 3¢-end formation, and their perturbation underlies several diseases. Recent research from my lab and others suggests that this group of ncRNAs is more functionally diverse than previously thought and that the canonical functions of snRNAs in splicing and 3¢-end processing constitute only the tip of the iceberg. Using Herpesvirus saimiri (HVS) as a model system, we recently showed that snRNAs perform functions beyond pre-mRNA processing. This primate g-herpesvirus expresses seven snRNAs in latently infected cells called HSURs (Herpesvirus saimiri U-rich RNAs). We have focused on two of these ncRNAs, HSURs 1 and 2, because they are the most highly conserved HSURs among different HVS groups and the only two expressed by the related Herpesvirus ateles. HSUR1 and HSUR2 basepair with host microRNAs (miRNAs) and target one miRNA, miR-27, for degradation. Work from our laboratory showed that HSUR2 also basepairs with host mRNAs and links bound miRNAs with specific mRNAs to destabilize them. This constitutes the first example of an snRNA function after pre-mRNA processing. We also developed iRICC, a technique to determine RNA binding partners of a single RNA of interest and the sequences mediating RNA-RNA interactions in vivo. Using iRICC we showed that HSUR2 binding sites reside mostly in the 3¢ untranslated regions (3¢UTRs) of target mRNAs. We also showed that HSUR2 does not present a “seed” or specialized region that is used to interact with most targets, but rather acts as a flexible adaptor that interacts through different base-pairing arrangements with different mRNAs. iRICC revealed that HSUR1 profusely binds to coding sequences in addition to 3¢UTRs of target mRNAs. These findings suggest that HSUR1 might regulate host gene expression through mechanisms different from those employed by HSUR2. Finally, we have also developed an approach for identifying low-abundance snRNAs and discovered that mammalian cells express previously uncharacterized snRNAs. We will use a combination of genetic, genome-wide, molecular biology and biochemical approaches to further characterize mechanical aspects of HSUR1 and HSUR2 function and to functionally characterize novel cellular snRNAs. This work will advance our understanding of the mechanisms underlying the regulation of gene expression by this class on ncRNAs during viral infection. It will also illuminate novel mechanisms of gene regulation in the broad range of eukaryotes that express snRNAs.

概括 我实验室的总体目标是定义一组非 - 的功能和机制 编码RNA（NCRNA）：小核，富含U的RNA（SNRNA）。细胞SNRNA与SM蛋白辅助 形成核糖核蛋白颗粒（RNP），是负责的细胞机械的重要组成部分 剪接和3¢末端形成的前体信使RNA（前MRNA）处理步骤及其 扰动是几种疾病的基础。我的实验室和其他人的最新研究表明 NCRNA在功能上比以前认为的更多样化，并且SNRNA的规范功能 剪接和3¢末端处理仅构成冰山一角。 使用疱疹病毒Saimiri（HVS）作为模型系统，我们最近表明SNRNA的功能 超越前MRNA处理。这个灵长类动物的G- Hepesvirus在潜在感染的细胞中表达七个SNRNA 称为基人（疱疹病毒saimiri u-rich RNA）。我们专注于这两个NCRNA，HSURS 1和2， 因为它们是不同HVS组中最高保守的HSUS，而仅有两个表示 由相关的疱疹病毒垫圈。 HSUR1和HSUR2 BASEPAIR，带有宿主microRNA（miRNA），靶向一个 miRNA，miR-27，用于降解。我们实验室的工作表明，HSUR2也带有宿主mRNA的基础 并将miRNA与特定的mRNA联系起来，以破坏它们的稳定。这构成了snRNA的第一个例子 MRNA处理后的功能。我们还开发了IRICC，这是一种确定RNA结合伙伴的技术 单个感兴趣的RNA和在体内介导RNA-RNA相互作用的序列。使用IRICC我们显示 HSUR2结合位点主要存在于目标mRNA的3¢未翻译区域（3¢UTR）。我们也是 表明HSUR2不存在用于与大多数目标相互作用的“种子”或专业区域 而是充当一个灵活的适配器，通过不同的基础配对安排与不同的相互作用 mrnas。 IRICC表明，HSUR1除了3¢utrs外，还与编码序列结合 mrnas。这些发现表明HSUR1可能通过不同的机制来调节宿主基因的表达 从HSUR2雇用的人。最后，我们还开发了一种方法来识别低收入的方法 SNRNA并发现哺乳动物细胞表达先前未表征的SNRNA。 我们将结合遗传，全基因组，分子生物学和生化方法的结合 进一步表征HSUR1和HSUR2功能的机械方面，并在功能上表征新颖 细胞snrnas。这项工作将促进我们对调节机制的理解 该类别在病毒感染期间对NCRNA的基因表达。它还将阐明 在表达SNRNA的广泛真核生物中的基因调节。