Transposition Mechanisms

换位机制

• 批准号: 8050573
• 负责人: Jef D BOEKE
• 金额: $ 51.48万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1986
• 资助国家: 美国
• 起止时间: 1986-04-01 至 2014-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8050573
• 关键词: Acquired Immunodeficiency Syndrome; Base Pairing; Beryllium; Biochemical; Biological Models; Cells; Complementary DNA; DNA; DNA Damage; DNA Polymerase III; DNA Structure; Defect; Elements; Enzymes; Eukaryota; Genes; Genetic; Genetic Transcription; Genome; Genome Stability; Genomics; Goals; HIV-1; Human Genome; In Vitro; Integrase; Integration Host Factors; Jumping Genes; Lead; Life Cycle Stages; Malignant Neoplasms; Maps; Mediating; Meiosis; Mitosis; Modeling; Molecular Genetics; Play; Positioning Attribute; Process; RNA; RNA Polymerase II; RNA Polymerase III; RNA Processing; RNA-Directed DNA Polymerase; Retroelements; Retrotransposition; Retrotransposon; Role; Time; Transfer RNA; Virus; Yeasts; design; human DNA; in vivo; insight; interest; lariat debranching enzyme; microbial; synthetic biology; tool

DESCRIPTION (provided by applicant): The retrotransposon Ty1 has served as a key model system for understanding the replication and integration of retroelements. In addition to providing key insights into replication mechanisms through ongoing mechanistic studies, it has become clear that Ty1 is an outstanding model for understanding the intimate relationship between retroelements and their host genomes. Replication of Ty1 elements is mediated by host RNA polymerase II, the Ty1-encoded reverse transcriptase (RT) enzyme, and a cellular tRNA primer, all acting on Ty1 RNA. Using a synthetic biology approach, we will produce a saturating map of the RNA and DNA structures required for retrotransposition. RNA apparently plays an as yet unclear role in retrotransposition, because Ty1 retrotransposition requires a several RNA processing enzymes including the RNA lariat debranching enzyme Dbr1. Integration of Ty1 cDNA is mediated by integrase (IN). Integration is targeted to very specific regions of the host genome, namely integration windows of several hundred base pairs immediately upstream of RNA polymerase III-transcribed genes. We are exploring this process through traditional genetic and biochemical studies as well as by using three new tools we have developed, 1) Design and analysis of synthetic Ty1 elements, 2) a Transposon Insertion Profiling chip (TIP-chip) and 3) ultra-high copy (UHC) Ty1 strains that contain more than 10 times the load of Ty1 elements carried by wild- type strains. The latter is an especially interesting microbial model for the relationship between retroelements and the genomes of higher eukaryotes, such as the human genome. A combined genetic, molecular and genomic approach will be taken to further dissect the Ty1 life cycle and Ty1's intimate relationship with its host genome.GM036481-A1 Project Narrative: The goal of this project is to deepen our understanding of how "jumping genes" change positions in a yeast cell's DNA. This mechanism is similar to the mechanism used by the HIV-1 virus (which causes AIDS) when it "jumps" into human DNA. Additionally, we study how the jumping genes interact with the machinery that protects our DNA from damage, a process that can lead to cancer.

描述（由申请人提供）：Retrotransposon Ty1已成为理解恢复元素的复制和集成的关键模型系统。除了通过持续的机械研究提供对复制机制的关键见解外，很明显，Ty1是理解逆向元素及其宿主基因组之间亲密关系的杰出模型。 TY1元件的复制是由宿主RNA聚合酶II，TY1编码的逆转录酶（RT）酶和细胞TRNA底漆介导的，所有这些酶都作用于TY1 RNA。使用合成生物学方法，我们将产生逆转录置所需的RNA和DNA结构的饱和图。 RNA显然在逆转录中起着尚不清楚的作用，因为TY1逆转录置次置位需要几种RNA加工酶，包括RNA Lariat脱粒酶DBR1。 TY1 cDNA的整合是由整合酶（IN）介导的。集成针对宿主基因组的非常特异性的区域，即几百个碱基对的整合窗口，立即在RNA聚合酶III转录基因的上游。 We are exploring this process through traditional genetic and biochemical studies as well as by using three new tools we have developed, 1) Design and analysis of synthetic Ty1 elements, 2) a Transposon Insertion Profiling chip (TIP-chip) and 3) ultra-high copy (UHC) Ty1 strains that contain more than 10 times the load of Ty1 elements carried by wild- type strains.后者是一个特别有趣的微生物模型，用于追溯元素与较高真核生物（例如人类基因组）的基因组之间的关系。将采用一种遗传，分子和基因组方法，以进一步剖析TY1生命周期和TY1与宿主基因组的亲密关系。GM036481-A1 项目叙述：该项目的目的是加深我们对“跳跃基因”如何改变酵母细胞DNA的位置的理解。该机制类似于HIV-1病毒（引起艾滋病）“跳入人类DNA”时使用的机制。此外，我们研究了跳跃基因如何与保护我们的DNA免受损害的机械相互作用，这一过程可能导致癌症。

项目成果

Transposon-mediated genome manipulation in vertebrates.

• DOI: 10.1038/nmeth.1332
• 发表时间: 2009-06
• 期刊: NATURE METHODS
• 影响因子: 48
• 作者: Ivics, Zoltan;Li, Meng Amy;Mates, Lajos;Boeke, Jef D.;Nagy, Andras;Bradley, Allan;Izsvak, Zsuzsanna
• 通讯作者: Izsvak, Zsuzsanna

A nucleocapsid functionality contained within the amino terminus of the Ty1 protease that is distinct and separable from proteolytic activity.

Ty1 蛋白酶氨基末端包含的核衣壳功能与蛋白水解活性不同且可分离。

• DOI: 10.1128/jvi.76.1.346-354.2002
• 发表时间: 2002
• 期刊: Journal of virology
• 影响因子: 5.4
• 作者: LawlerJr,JosephF;Merkulov,GennadyV;Boeke,JefD
• 通讯作者: Boeke,JefD

High-frequency deletion between homologous sequences during retrotransposition of Ty elements in Saccharomyces cerevisiae.

酿酒酵母 Ty 元件逆转座过程中同源序列之间的高频缺失。

• DOI: 10.1073/pnas.84.23.8553
• 发表时间: 1987
• 期刊: Proceedings of the National Academy of Sciences of the United States of America
• 影响因子: 11.1
• 作者: Xu,H;Boeke,JD
• 通讯作者: Boeke,JD

In-frame linker insertion mutagenesis of yeast transposon Ty1: mutations, transposition and dominance.

酵母转座子 Ty1 的框内接头插入诱变：突变、转座和显性。

• DOI: 10.1016/0378-1119(94)90517-7
• 发表时间: 1994
• 期刊: Gene
• 影响因子: 3.5
• 作者: Monokian,GM;Braiterman,LT;Boeke,JD
• 通讯作者: Boeke,JD

In vitro integration of retrotransposon Ty1: a direct physical assay.

逆转录转座子 Ty1 的体外整合：直接物理测定。

• DOI: 10.1128/mcb.14.9.5719-5730.1994
• 发表时间: 1994
• 期刊: Molecular and cellular biology
• 影响因子: 5.3
• 作者: Braiterman,LT;Boeke,JD
• 通讯作者: Boeke,JD