The molecular characterization of sex-specific piRNA transcription and snRNA transcription in C. elegans

线虫性别特异性 piRNA 转录和 snRNA 转录的分子特征

• 批准号: 10464652
• 负责人: Lars Benner
• 金额: $ 4.68万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-06-01 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10464652
• 关键词: Address; Adult; Aging; Alzheimer' s Disease; Amino Acid Motifs; Amino Acid Sequence; Animal Model; Animals; Binding; Binding Proteins; Binding Sites; Biogenesis; CRISPR/Cas technology; Caenorhabditis elegans; Complement; Complex; DNA Binding; DNA Transposable Elements; DNA-Binding Proteins; Data; Disease; Drosophila melanogaster; Exhibits; Exonuclease; Family; Female; Fertility; Fertility Disorders; Gene Targeting; Genetic Transcription; Genomic Instability; Genomics; Germ Cells; Global Change; Human; Hybrids; Infertility; Knowledge; Link; Male Infertility; Malignant Neoplasms; Mammals; Mediating; Molecular; Mus; Nematoda; Neurons; Organism; Orthologous Gene; Pathway interactions; Play; Protein Family; Proteins; RNA; RNA Interference; RNA Splicing; Reagent; Role; Small Nuclear RNA; Small RNA; Specific qualifier value; Specificity; Spliceosomes; Sterility; Tertiary Protein Structure; Testis; Work; Zebrafish; base; experimental study; fly; genome integrity; genomic RNA; genomic locus; insight; male; mutant; offspring; paralogous gene; preservation; protein complex; recruit; sex; sexual dimorphism; transcription factor; transcriptome sequencing

Project Summary The molecular characterization of sex-specific piRNA transcription and snRNA transcription in C. elegans. PIWI-interacting RNAs (piRNAs) play key roles in repressing transposons in the germline. Loss of transposon- targeting piRNAs can have sex-specific consequences leading to genomic instability and infertility. However, the mechanisms of sex-specific piRNA expression remain largely unknown. Recent studies have revealed that SNPC-4, a core factor required for snRNA transcription, is also essential for piRNA transcription in C. elegans and suggests that the snRNA transcriptional machinery may have been co-opted to transcribe piRNAs. Therefore, identifying the transcription factors and characterizing the specific protein domains that dictate sex- specific piRNA and snRNA transcriptional specificity will address this critical knowledge gap. The small nuclear RNA activating protein complex (SNAPc) is a conserved heterotrimeric complex consisting of SNAPC1, SNAPC3, and SNAPC4 that facilitates snRNA transcription in mammals. Unlike mammals, C. elegans have several orthologs of SNAPC1 that have sexually dimorphic roles in piRNA biogenesis. My preliminary data show that SNPC-1.2 promotes female piRNA transcription while SNPC-1.3 is a known male piRNA transcription factor. Furthermore, my initial data show the C. elegans SNAPC3 ortholog, SNPC-3.4, facilitates snRNA biogenesis, while SNPC-3.1 and SNPC-3.2 act redundantly to drive both male and female piRNA expression. I will investigate my hypothesis that SNPC-1.2 is a bona fide female piRNA transcription factor and harbors unique protein motifs conferring female piRNA expression, while distinct domains in SNPC-1.3 specify male piRNA expression (Aim 1). Additionally, I predict SNPC-3.4 is a snRNA biogenesis factor, while SNPC-3.1 and SNPC-3.2 act redundantly to drive piRNA transcription. Analogous to the SNPC1 family, I predict that distinct domains within SNPC-3.4 and SNPC-3.1/3.2 specify their recruitment and transcription at snRNA and piRNA genomic loci, respectively (Aim 2). Collectively, these data will elucidate the snRNA and female and male piRNA transcriptional complexes and identify the protein motifs that engender snRNA and sex-specific piRNA transcription. This work may have implications in treating fertility defects that are due to abnormal piRNA expression as well as different neuronal diseases associated with snRNA misregulation such as ALS.

项目摘要 秀丽隐杆线虫中性别特异性皮肤转录和snRNA转录的分子表征。 Piwi-Interacting RNA（PIRNA）在抑制种系中的转座子中起关键作用。转座子的损失 靶向PIRNA可能会导致基因组不稳定性和不育症。但是， 性别特异性piRNA表达的机制在很大程度上未知。最近的研究表明 SNPC-4是SnRNA转录所需的核心因素，对于秀丽隐杆线虫中的piRNA转录也是必不可少的 并表明SNRNA转录机械可能已选择转录PIRNA。 因此，确定转录因子并表征决定性别的特定蛋白质结构域 特定的PIRNA和SNRNA的转录特异性将解决这一关键知识差距。小核 RNA激活蛋白质复合物（SNAPC）是由SNAPC1组成的保守异三聚体复合物 SNAPC3和SNAPC4促进哺乳动物中的SnRNA转录。与哺乳动物不同，秀丽隐杆线虫有 SNAPC1的几种直系同源物在PIRNA生物发生中具有性二态作用。我的初步数据显示 SNPC-1.2促进雌性piRNA转录，而SNPC-1.3是已知的雄性皮纳转录因子。 此外，我的初始数据显示了秀丽隐杆线虫SNAPC3直系同源物，SNPC-3.4，促进了SnRNA生物发生， 而SNPC-3.1和SNPC-3.2起作用，以驱动雄性和女性胸肌的表达。我会调查 我的假设是SNPC-1.2是一个真正的女性胸肌转录因子，并且拥有独特的蛋白质基序 赋予雌性piRNA的表达，而SNPC-1.3中的不同域则指定雄性piRNA表达（AIM 1）。此外，我预测SNPC-3.4是SNRNA生物发生因子，而SNPC-3.1和SNPC-3.2冗余地起作用。 驱动Pirna转录。与SNPC1家族相似，我预测SNPC-3.4中的不同域 SNPC-3.1/3.2分别指定其在SnRNA和PIRNA基因组基因座的募集和转录 （目标2）。总的来说，这些数据将阐明snRNA以及女性和雄性PIRNA转录复合物 并确定引起snRNA和性别特异性piRNA转录的蛋白质基序。这项工作可能有 在治疗piRNA异常表达以及不同神经元引起的生育缺陷中的含义 与SNRNA失调有关的疾病，例如ALS。