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

基本信息

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
项目类别：
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&apos 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

项目摘要

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.

项目概要线虫中性别特异性 piRNA 转录和 snRNA 转录的分子特征。 PIWI 相互作用 RNA (piRNA) 在抑制种系转座子方面发挥着关键作用。转座子丢失- 靶向 piRNA 可能会产生性别特异性后果，导致基因组不稳定和不育。然而，性别特异性 piRNA 表达的机制仍然很大程度上未知。最近的研究表明 SNPC-4 是 snRNA 转录所需的核心因子，也是线虫中 piRNA 转录所必需的并表明 snRNA 转录机制可能被用来转录 piRNA。因此，鉴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 转录特异性将解决这一关键的知识空白。小核 RNA 激活蛋白复合物 (SNAPc) 是一种保守的异三聚体复合物，由 SNAPC1、 SNAPC3 和 SNAPC4 促进哺乳动物中 snRNA 的转录。与哺乳动物不同，秀丽隐杆线虫 SNAPC1 的几种直系同源物在 piRNA 生物发生中具有性别二态性作用。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 piRNA 转录因子。此外，我的初始数据显示线虫 SNAPC3 直向同源物 SNPC-3.4 促进 snRNA 生物发生，而SNPC-3.1和SNPC-3.2则冗余地驱动雄性和雌性piRNA表达。我会调查我的假设是 SNPC-1.2 是一个真正的雌性 piRNA 转录因子并具有独特的蛋白质基序赋予雌性 piRNA 表达，而 SNPC-1.3 中的不同结构域指定雄性 piRNA 表达（目的 1）。此外，我预测 SNPC-3.4 是一个 snRNA 生物发生因子，而 SNPC-3.1 和 SNPC-3.2 的作用是多余的驱动 piRNA 转录。与 SNPC1 家族类似，我预测 SNPC-3.4 中的不同域和 SNPC-3.1/3.2 分别指定它们在 snRNA 和 piRNA 基因组位点的招募和转录（目标 2）。总的来说，这些数据将阐明 snRNA 以及雌性和雄性 piRNA 转录复合物并鉴定产生 snRNA 和性别特异性 piRNA 转录的蛋白质基序。这部作品可能有治疗由于 piRNA 表达异常以及不同神经元导致的生育缺陷的影响与 snRNA 失调相关的疾病，例如 ALS。