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Retrotransposon expression within ribosomal gene loci

核糖体基因座内的逆转录转座子表达

基本信息

批准号：
8396384
负责人：
Thomas H. Eickbush
金额：
$ 43.04万
依托单位：
UNIVERSITY OF ROCHESTER
依托单位国家：
美国
项目类别：
财政年份：
1992
资助国家：
美国
起止时间：
1992-08-01 至 2014-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8396384
关键词：
Affect Amino Acid Motifs Amino Acid Substitution Animals Architecture Beryllium Binding Biochemical Biological Models Cell Nucleolus Cells Chromatin Chromosome Structures DNA DNA Transposable Elements Dosage Compensation (Genetics)Drosophila genus Elements Epigenetic Process Evolution Family Gene Expression Regulation Genes Genetic Recombination Genetic Transcription Genome Goals Growth Hereditary Disease Human Human Genome In Vitro Indium Introns Investigation Laboratories Lead Link Malignant Neoplasms Metabolism Methods Modeling Monitor Morphology Mutation Onset of illness Organism Pathway interactions Play Population Positioning Attribute Protein Binding Proteins RNA RNA Binding RNA Interference RNA Processing Reaction Recombinant DNA Regulation Research Retrotransposition Retrotransposon Reverse Transcription Ribosomal RNA Role Series Shapes Signal Pathway Site Small RNA Specificity Structure System Telomerase Testing Time Transcript Transcriptional Regulation Work Y Chromosome base design insight male public health relevance rRNA Genes research study

项目摘要

DESCRIPTION (provided by applicant): Only a small fraction of eukaryotic genomes encode protein or is involved in the regulation of gene expression. A far larger fraction of these genomes has little or no function and is derived from the reverse transcription of RNA. For example, at least 40% of the human genome is composed of these reverse transcripts. The gradual accumulation of such insertions over time has played a significant role in shaping the size, structure and function of our genome. The family of retrotransposable elements known as LINEs generates the protein machinery responsible for most of these insertions. One of the best model systems in which to study LINEs is R2, an element that inserts in a sequence specific manner into a fraction of the hundreds of tandemly repeated 28S rRNA genes found in all higher organisms. The high degree of sequence specificity of the R2 integration reaction has enabled detailed biochemical studies of its mechanism. A critical but poorly understood aspect of this integration mechanism is how the R2 protein binds the two ends of the R2 RNA template used for reverse transcription. One series of experiments in this proposal is a detailed study of amino acid substitutions in the R2 protein that influence its ability to bind these RNA regions. These studies will contribute to a better understanding of how LINE elements are responsible for many of the insertions that occur in a more random manner throughout the human genome. Another goal of this proposal is to understand how R2 elements are regulated. The tandemly repeated rRNA genes (rDNA locus) form the nucleolus, the site of rRNA synthesis and ribosomal subunit assembly. While each R2 insertion disrupts the function of one 28S rRNA gene, an organism can survive as long as sufficient numbers of rRNA genes remain uninserted. Thus a second series of experiments is designed to determine how organisms are able to generate high levels of rRNA from uninserted genes while minimizing the expression of R2 elements from the otherwise identical inserted rRNA genes. Transcription of inserted rRNA genes gives rise to new R2 insertion. Using methods to monitor the transcription of specific R2 elements and to position these elements within the rDNA locus, the regions of the rDNA locus that are transcribed will be defined, and these transcribed regions monitored over time to determine how they are influenced by recombination and R2 element activity. Finally, mutations in a number of genes involved in chromosome structure and rRNA gene regulation will be tested in a third series of experiments to determine whether they affect the ability of the organism to differentiate between inserted and uninserted 28S genes. These experiments should reveal new insights into how transcription of the entire rDNA locus and its R2 elements are differentially regulated.

描述（由申请人提供）：只有一小部分真核基因组编码蛋白质或参与基因表达调控。这些基因组中有很大一部分几乎没有或没有功能，并且来自RNA的逆转录。例如，至少40%的人类基因组由这些反向转录物组成。随着时间的推移，这种插入的逐渐积累在塑造我们基因组的大小，结构和功能方面发挥了重要作用。逆转录转座因子家族（称为LINE）产生负责大多数这些插入的蛋白质机制。研究LINES的最佳模型系统之一是R2，它是一种以序列特异性方式插入所有高等生物中发现的数百个串联重复的28S rRNA基因中的一小部分的元件。R2整合反应的高度序列特异性使得能够对其机制进行详细的生物化学研究。这种整合机制的一个关键但知之甚少的方面是R2蛋白如何结合用于逆转录的R2 RNA模板的两端。该提案中的一系列实验是详细研究R2蛋白中影响其结合这些RNA区域的能力的氨基酸取代。这些研究将有助于更好地理解LINE元件如何负责在整个人类基因组中以更随机的方式发生的许多插入。该提案的另一个目标是了解R2元件是如何调节的。串联重复的rRNA基因（rDNA基因座）形成核仁，即rRNA合成和核糖体亚基组装的场所。虽然每个R2插入会破坏一个28S rRNA基因的功能，但只要有足够数量的rRNA基因未插入，生物体就可以存活。因此，第二系列的实验被设计来确定生物体如何能够从未插入的基因产生高水平的rRNA，同时最小化来自在其他方面相同的插入的rRNA基因的R2元件的表达。插入的rRNA基因的转录产生新的R2插入。使用监测特定R2元件的转录并将这些元件定位在rDNA基因座内的方法，将定义rDNA基因座的转录区域，并随时间监测这些转录区域以确定它们如何受到重组和R2元件活性的影响。最后，将在第三系列实验中测试涉及染色体结构和rRNA基因调控的许多基因的突变，以确定它们是否影响生物体区分插入和未插入的28S基因的能力。这些实验应该揭示新的见解如何转录的整个rDNA基因座和它的R2元件的差异调节。

项目成果

期刊论文数量（19）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Preferential occupancy of R2 retroelements on the B chromosomes of the grasshopper Eyprepocnemis plorans.

DOI：
10.1371/journal.pone.0091820
发表时间：
2014
期刊：
PloS one
影响因子：
3.7
作者：
Montiel EE;Cabrero J;Ruiz-Estévez M;Burke WD;Eickbush TH;Camacho JP;López-León MD
通讯作者：
López-León MD

The pattern of R2 retrotransposon activity in natural populations of Drosophila simulans reflects the dynamic nature of the rDNA locus.

DOI：
10.1371/journal.pgen.1000386
发表时间：
2009-02
期刊：
PLoS genetics
影响因子：
4.5
作者：
Zhou J;Eickbush TH
通讯作者：
Eickbush TH

The reverse transcriptase encoded by the non-LTR retrotransposon R2 is as error-prone as that encoded by HIV-1.

非 LTR 逆转录转座子 R2 编码的逆转录酶与 HIV-1 编码的逆转录酶一样容易出错。

DOI：
10.1016/j.jmb.2011.02.015
发表时间：
2011
期刊：
Journal of molecular biology
影响因子：
5.6
作者：
Jamburuthugoda,VaruniK;Eickbush,ThomasH
通讯作者：
Eickbush,ThomasH

The R2 retrotransposon RNA families.

R2 逆转录转座子 RNA 家族。

DOI：
10.4161/rna.8.5.16033
发表时间：
2011
期刊：
RNA biology
影响因子：
4.1
作者：
Moss,WalterN;Eickbush,DannaG;Lopez,MichaelJ;Eickbush,ThomasH;Turner,DouglasH
通讯作者：
Turner,DouglasH

Evolution of the R2 retrotransposon ribozyme and its self-cleavage site.