Transposition of Tn7 and Hermes

Tn7 和 Hermes 的转位

• 批准号: 7577358
• 负责人: NANCY Craig CRAIG
• 金额: $ 31.05万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-03-01 至 2012-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7577358
• 关键词: Acyl Carrier Protein; Antibiotic Resistance; Antigen Receptors; Binding; Biochemical; Biological Assay; Cells; Chromosomal Rearrangement; Chromosomal translocation; Complex; DNA; DNA Damage; DNA Transposable Elements; Dissection; Double-Stranded RNA; Drosophila genus; Drosophila melanogaster; Elements; Escherichia coli; Event; Excision; Frequencies; Gene Expression; Genes; Genetic; Genetic Recombination; Genome; Genomic Instability; Human Genome; In Vitro; Individual; Insecta; Integration Host Factors; Lead; Libraries; Malignant - descriptor; Mediating; Methodology; Molecular; Molecular Chaperones; Movement; Nonhomologous DNA End Joining; Nucleoproteins; Oncogenic; Organism; Paste substance; Peptide Hydrolases; Plasmids; Positioning Attribute; Process; Property; Proteins; RNA Interference; Reaction; Receptor Gene; Relative (related person); Research Personnel; Resolution; Role; Saccharomyces cerevisiae; Screening procedure; Side; Signal Transduction; Site; System; Transposase; V(D)J Recombination; Visual; Work; Yeasts; fly; in vivo; interest; lymphoid neoplasm; mutant; programs; recombinase; repaired; research study; ribosomal protein L29

Although the constancy of genetic information is central, genomes are surprisingly unstable. Transposable elements, i.e.discrete DNA segments that can move between many non-homologous positions, are widespread, having been found in virtually all organisms. Indeed, in some genomes, transposable element sequences are the predominantcomponent: strikingly, about 45% of the human genome is composed of transposable element sequences. The consequences of transposable element movement are many: insertion can result in alterations in host gene expression, in the covalent linkage of element-encoded determinates to host DNA, for example the joining of antibiotic resistance determinants to plasmids that can move between cells or the integration of retroviral DNA into host genomes. Also, gaps in the donor DNA followingelement excision are often not restored to their pre- transposon state. Moreover, DNA breaks can result in DNA damage signals. We propose to study at the molecular and biochemical level how transposition is controlled by its host and how the host responds to the movement of transposable elements, a potentially lethal event because it involves DNA breakage and joining. We will study the bacterial transposon Tn7 in E. coli and the insect transposon Hermes in S. cerevisiae and in Drosophila. We will probe how the assembly of particular nucleoprotein in complexes can control transposition though in vitro dissection of Tn7 transposition and will dissect the role of host functions in Tn7 transposition through the isolation and characterization of host mutants that alter transposition. We will also probe Hermes transposition in yeast and flies through the isolation of mutants that alter its movement. We will also examine how the transposase-mediated breaks can lead to chromosomal rearrangements, i.e. genetic instability.

尽管遗传信息的恒定是中心的，但基因组令人惊讶地不稳定。 转座元素，即可以在许多非同源物之间移动的dncrete DNA段 在几乎所有生物体中都发现了位置，这是广泛的。确实，在某些基因组中， 转座元素序列是主要组成部分：令人惊讶的是，大约45％的人 基因组由转座元序列组成。可转座元素的后果 运动很多：插入可以导致宿主基因表达的改变，在共价链接中 元素编码的确定宿主DNA的确定，例如抗生素耐药性的连接 可以在细胞之间移动或将逆转录病毒DNA整合到宿主之间移动的质粒的决定因素 基因组。同样，供体DNA后插入切除的差距通常不会恢复到其前 跨盆地状态。此外，DNA断裂会导致DNA损伤信号。我们建议在 分子和生化水平如何由宿主控制换位以及宿主如何控制 响应可转移元素的运动，这是一个潜在的致命事件，因为它涉及 DNA断裂和加入。我们将研究大肠杆菌和昆虫的细菌转座子TN7 在酿酒酵母和果蝇中的转座爱马仕。我们将探测特定的组装 通过在TN7转座的体外解剖，复合物中的核蛋白可以控制转座 并将通过隔离和 改变换位的宿主突变体的表征。我们还将探测爱马仕的换位 通过隔离突变体改变其运动的酵母。我们还将研究如何 转座酶介导的断裂可以导致染色体重排，即遗传不稳定性。