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: 10590605
• 负责人: Lars Benner
• 金额: $ 4.77万
• 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/10590605
• 关键词: Address; Adult; Aging; Alzheimer' s Disease; Amino Acid Motifs; Amino Acid Sequence; 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 Splicing; Reagent; Repression; Role; Small Nuclear RNA; Small RNA; Specific qualifier value; Specificity; Spliceosomes; Sterility; Tertiary Protein Structure; Testis; Work; Zebrafish; experimental study; fly; genome integrity; genomic locus; insight; male; model organism; mutant; offspring; paralogous gene; piRNA; 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 对C.优美的 PIWI相互作用RNA（piRNA）在生殖系中抑制转座子中起关键作用。转座子丢失- 靶向piRNA可具有性别特异性后果，导致基因组不稳定性和不育。但 性别特异性皮尔纳表达的机制在很大程度上仍然未知。最近的研究表明， SNPC-4是皮尔纳转录所需的核心因子，也是C. elegans 并表明snRNA转录机制可能已被增选以转录piRNA。 因此，确定转录因子和表征决定性别的特定蛋白质结构域- 特异性皮尔纳和snRNA转录特异性将解决这一关键的知识缺口。小核 RNA激活蛋白复合物（SNAPc）是一种保守的异源三聚体复合物，由SNAPC 1、 SNAPC 3和SNAPC 4，其促进哺乳动物中的snRNA转录。与哺乳动物不同，C.优雅的有 SNAPC 1的几种直系同源物在皮尔纳生物发生中具有性二态性作用。我的初步数据显示 SNPC-1.2促进雌性皮尔纳转录，而SNPC-1.3是已知的雄性皮尔纳转录因子。 此外，我的初步数据显示，C。线虫SNAPC 3直系同源物SNPC-3.4，促进snRNA生物合成， 而SNPC-3.1和SNPC-3.2冗余地驱动雄性和雌性皮尔纳表达。我会调查 我的假设是SNPC-1.2是一个真正的女性皮尔纳转录因子，具有独特的蛋白质基序， 赋予雌性皮尔纳表达，而SNPC-1.3中的不同结构域指定雄性皮尔纳表达（目的 1）。另外，我预测SNPC-3.4是一个snRNA生物合成因子，而SNPC-3.1和SNPC-3.2是冗余的 来驱动皮尔纳转录。类似于SNPC 1家族，我预测SNPC-3.4中的不同结构域 和SNPC-3.1/3.2分别在snRNA和皮尔纳基因组基因座上指定它们的募集和转录 (Aim 2）。总的来说，这些数据将阐明snRNA和雌性和雄性皮尔纳转录复合物 并鉴定产生snRNA和性别特异性皮尔纳转录的蛋白基序。这项工作可能有 在治疗由于异常皮尔纳表达以及不同的神经元细胞引起的生育缺陷方面的意义 与snRNA失调相关的疾病，如ALS。