Determinants of SINE mobilization in the domestic dog.

家犬正弦动员的决定因素。

基本信息

批准号：
10046792
负责人：
Julia Vera Halo
金额：
$ 42.6万
依托单位：
BOWLING GREEN STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-09-01 至 2023-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10046792
关键词：
7SL RNA Address Affect Alu Elements Animal Model Atrophic Binding Biochemical Biological Biological Assay Biological Models Body Size Canis familiaris Centronuclear myopathy Color Comparative Genomic Analysis Complement Consensus Conserved Sequence DNA Polymerase III DNA Transposable Elements Data Disease Docking Elements Engineering Event Evolution Exhibits Experimental Genetics Exposure to Family Foundations Future Gene Frequency Genetic Polymorphism Genetic Variation Genome Genomics Genotype Goals Health Human Human Biology Individual Insertional Mutagenesis Integration Host Factors Lead Link Locales Long Interspersed Elements Lysine-Specific tRNA Malignant Neoplasms Mammals Mediator of activation protein Messenger RNA Methods Minority Modeling Molecular Genetics Mutagenesis Mutation Narcolepsy ORF2 protein Orthologous Gene Outcome Paste substance Pathway interactions Pattern Phenotype Plasmids Poly A Poly(A) Tail Population Process Property Proteins Publishing RNA Research Resources Retina Retrotransposition Ribonucleoproteins Ribosomes Role Sequence Analysis Short Interspersed Nucleotide Elements Signal Recognition Particle Site Structure Transfer RNA Translations Variant Work base comparative genome wide association study genomic variation homologous recombination human model improved interdisciplinary approach member monomer protein function recruit tissue culture

项目摘要

Abstract Transposable elements (TEs) have abundantly colonized eukaryotic genomes and are profound mediators of genomic variation, phenotypes, and disease. In humans and other mammals, the vast majority of TE families are defective due to accumulated mutations within the host, but some remain capable of generating new copies. Intracellular spread of such elements can lead to sporadic disease and variation via insertional mutagenesis or by facilitating rearrangements through non-allelic homologous recombination. Understanding the mutagenic potential offered from a particular TE family requires refinement of its lineages, individual copies, and functional properties that affect mobilization capacities within the host. The short and long interspersed elements (SINEs and LINEs) mobilize through an intracellular ‘copy and paste’ process termed retrotransposition. SINEs are non- autonomous and mobilize by recruiting functions in trans from a retrotransposition-competent LINE, accomplished in part by L1 ORF2 protein (ORF2p) poly(A)-tail recognition on the RNA intermediate. The human SINE Alu originated ~65 mya from the 7SL RNA of signal recognition particle (SRP), a cytoplasmic ribonucleoprotein that docks on the ribosome during co-translational targeting to the secretory pathway. The derived Alu exists as two 7SL ‘Alu’ monomers in tandem; both retain 7SL ancestral structure and cytoplasmic binding partners SRP9/14 that direct Alu RNA to ribosome, where it then diverts L1 ORF2p for its own encoding mRNA. Distinct in their origin, SINEs of tRNA ancestry include young families of conserved structure that are highly mobilized in some species. These observations and our own findings lead us to hypothesize that mobilization of the tRNA SINE intermediate is achieved by association with evolutionarily conserved cellular factors in a similar but biologically distinct model to that of Alu. We previously examined a mobile ~180bp SINE of tRNALys ancestry from the canine, SINCEC_Cf, that exhibits high levels of polymorphism, sequence conservation, and insertions linked to disease and artificially selected phenotypes. We have demonstrated bona fide L1Cf-driven retrotransposition of its own mRNA in cis and a synthetic SINCEC_Cf consensus in trans at rates considered ‘hot’ in comparison to those observed for Alu/L1Hs. Moreover, mobilization assays of naturally occurring SINCEC_Cf variants expose alterations to the consensus primary sequence that directly affect mobilization. We take advantage of this model to address our hypothesis and identify structural factors required for mobilization of the tRNA SINE using a combination of genomic, biochemical, molecular, and genetic experimental approaches. This proposal will provide the foundation for SINE mobilization properties in the canine model and a variant resource for comparative genomic analysis in application to human health.

抽象的转座元素（TES）具有丰富的真核基因组，是深刻的介体基因组变异，表型和疾病。由于宿主的累积突变Muthin而导致有缺陷，但有些仍然能够产生新副本。此类元素的细胞内扩散可以通过插入诱变或通过促进通过非平行性同源重组的重排。特定家族提供的潜力需要改进的是Linese，个体副本和功能影响主机中动员能力的特性。线条通过称为逆转的细胞内“复制和糊状”过程动员。自主和动员通过从转向倾倒竞争的线中募集功能，部分由L1 ORF2蛋白（ORF2P）poly（a）tail识别在RNA国际上正弦alu起源于〜65 Mya，来自信号识别粒子（SRP）的7SL RNA，一个细胞质核糖蛋白在分泌途径的核糖体上停靠派生的Alu作为两个7sl单体的存在；绑定伴侣SRP9/14将Alu引向核糖 mRNA。在某些物种中高度动员。 tRNA正弦中间体的动员通过与进化保守的细胞相关与Alu相似但生物学上不同的因素。犬Sincec_cf的Trnalys Ancestry的祖先，表现出高水平的多态性，序列保护以及与疾病和人工选择的表型有关的铭文。 fide l1CF驱动的逆转录在顺式中的mRNA和在at at at trans中的合成sincec_cf共识与ALU/L1H的观察到的速率II相比发生的SINCEC_CF变体暴露于直接影响共识的主要序列的变化动员。我们利用模型来解决我们的假设并确定所需的结构因素使用基因组，生化，分子和遗传的组合来动员tRNA正弦实验方法。模型和一种用于对人类健康应用的比较基因组分析的变体资源。