Genetic and Molecular Dissection of Meiotic Silencing by Unpaired DNA

不配对 DNA 减数分裂沉默的遗传和分子解析

• 批准号: 1715534
• 负责人: Patrick Shiu
• 金额: $ 69.96万
• 依托单位: University of Missouri-Columbia
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-08-01 至 2023-07-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1715534&HistoricalAwards=false
• 关键词: Genetic; Molecular; Dissection; Meiotic; Silencing

The goal of this project is to determine the underlying mechanism for how an unpaired DNA is silenced during meiosis, a process that produces gametes or spores. The presence of unpaired genes during this part of the cell cycle is often indicative of a foreign DNA element. Accordingly, the cell usually benefits by silencing (turning off) the expression of these genes. This pathway is called 'Meiotic Silencing by Unpaired DNA (MSUD)' and is a unique RNA-based gene silencing system first described in the orange bread mold, Neurospora crassa. But since DNA segments lacking a pairing partner are also targeted for silencing in worms, mice, and humans, the study of MSUD will shed light on how chromosome pairing affects gene regulation in higher organisms as well. To broaden the participation of underrepresented groups, the investigators of this project will recruit and train undergraduate students from their own university as well as from non-Ph.D.-granting and minority-serving institutions. Research activities from this project will be used to develop lesson plans for various outreach programs. Efforts will also be made to promote interdisciplinary partnerships and STEM (science, technology, engineering, and mathematics) education. In various organisms, genome surveillance systems are maintained to control the expression of repetitive elements and limit their spreading. One of these mechanisms is MSUD, which examines the pairing of any DNA segment with its homologous partner during meiotic prophase I. A gene not paired with a similar sequence generates a silencing signal (a small RNA species), which subsequently turns off the expression from this unpaired DNA as well as from any other (paired or unpaired) copies in the genome. MSUD utilizes common RNA interference (RNAi) factors (e.g., Argonaute) to target specific mRNA transcripts. Using the tools they have developed (e.g., protein interaction assays, mutant strains, RNA profiles, and cellular markers) and various complementary approaches (e.g., biochemistry, cytology, genetics, and molecular biology), the investigators plan to characterize Neurospora mutants defective in MSUD and examine how proteins from various sites of MSUD activity contribute to the silencing pathway.

这个项目的目标是确定未配对的DNA在减数分裂过程中如何沉默的潜在机制，减数分裂是产生配子或孢子的过程。在细胞周期的这一部分，未配对基因的存在通常表明存在外源DNA成分。因此，细胞通常通过沉默(关闭)这些基因的表达而受益。这一途径被称为不配对DNA减数分裂沉默(MSUD)，是一种独特的基于RNA的基因沉默系统，最初在橙色面包霉菌中被描述。但由于缺乏配对伴侣的DNA片段也是蠕虫、小鼠和人类沉默的目标，对MSUD的研究将揭示染色体配对如何影响高等生物体的基因调控。为了扩大代表不足群体的参与，该项目的调查人员将从他们自己的大学以及从非博士和少数群体服务的机构招聘和培训本科生。这个项目的研究活动将被用来为各种外展项目制定教案。还将努力促进跨学科伙伴关系和STEM(科学、技术、工程和数学)教育。在各种生物中，基因组监测系统被用来控制重复元件的表达并限制它们的传播。其中一种机制是MSUD，它在减数分裂前期I阶段检查任何DNA片段与其同源伙伴的配对。未与类似序列配对的基因会产生沉默信号(小RNA物种)，从而关闭该未配对DNA以及基因组中任何其他(配对或未配对)拷贝的表达。MSUD利用共同的RNA干扰(RNAi)因子(例如ArgAerte)来靶向特定的mRNA转录本。使用他们开发的工具(例如，蛋白质相互作用分析、突变菌株、RNA图谱和细胞标记)和各种互补方法(例如，生物化学、细胞学、遗传学和分子生物学)，研究人员计划表征MSUD中缺陷的脉孢子菌突变株，并研究MSUD活动的不同部位的蛋白质如何对沉默途径做出贡献。

项目成果

The Non-Coding RNA Journal Club: Highlights on Recent Papers—7

• DOI: 10.3390/ncrna5020040
• 发表时间: 2019-06
• 期刊: Non-Coding RNA
• 影响因子: 4.3
• 作者: Hua Xiao;P. Shiu;Marta Gabryleska;S. Conn;A. Dey;K. Chakrabarti;Manuel Regouc;M. Pichler;U. A. Ørom;G. Santulli;S. Nishiwada;A. Goel;Vaishnavi Nagarajan;L. Timmons;S. Alahari;N. Laprovitera;M. Ferracin;Po Hu;Hailing Jin

Identification of rfk-1, a Meiotic Driver Undergoing RNA Editing in Neurospora.

鉴定脉孢菌中正在进行 RNA 编辑的减数分裂驱动因子 rfk-1。

• DOI: 10.1534/genetics.119.302122
• 发表时间: 2019
• 期刊: Genetics
• 影响因子: 3.3
• 作者: Rhoades,NicholasA;Harvey,AustinM;Samarajeewa,DiliniA;Svedberg,Jesper;Yusifov,Aykhan;Abusharekh,Anna;Manitchotpisit,Pennapa;Brown,DarenW;Sharp,KevinJ;Rehard,DavidG;Peters,Joshua;Ostolaza-Maldonado,Xavier;Stephenson,Jackson;S
• 通讯作者: S

An RNA Recognition Motif-Containing Protein Functions in Meiotic Silencing by Unpaired DNA.

• DOI: 10.1534/g3.117.041848
• 发表时间: 2017-08-07
• 期刊: G3 (Bethesda, Md.)
• 作者: Samarajeewa DA;Manitchotpisit P;Henderson M;Xiao H;Rehard DG;Edwards KA;Shiu PKT;Hammond TM
• 通讯作者: Hammond TM

Suppressors of Meiotic Silencing by Unpaired DNA

不配对 DNA 减数分裂沉默的抑制剂

• DOI: 10.3390/ncrna5010014
• 发表时间: 2019
• 期刊: Non-Coding RNA
• 影响因子: 4.3
• 作者: Xiao, Hua;Hammond, Thomas;Shiu, Patrick
• 通讯作者: Shiu, Patrick