The function of germ granules in maintaining pluripotency in the C. elegans germline

胚芽颗粒在维持线虫种系多能性中的功能

基本信息

批准号：
10597973
负责人：
Dustin Lynn Updike
金额：
$ 33.2万
依托单位：
MOUNT DESERT ISLAND BIOLOGICAL LAB
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-04-01 至 2024-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10597973
关键词：
26S proteasome Affinity Alleles Animals Antigens Back Biogenesis CRISPR/Cas technology Caenorhabditis elegans Cataract Cell Nucleus Complex Core Protein Cytoplasm Cytoplasmic Granules DNA Data Development Elements Ensure Environment Epigenetic Process Equilibrium Exclusion Fertility Fertility Disorders Gene Expression Generations Genes Genetic Transcription Germ Germ Cells Goals Heritability Homologous Gene Human Individual Licensing Malignant Neoplasms Maps Mediating Messenger RNA Modeling Molecular Mutation Names Natural regeneration Nematoda Nuclear Nuclear Pore Peptide Initiation Factors Phase Polyribosomes Proteins Proteomics RNA RNA Binding RNA Helicase Reporter Reporting Research Ribonucleoproteins Ribosomes Role Safety Small RNA Somatic Cell Specificity Spermatogenesis Sterility Tertiary Protein Structure Testing Testis Time Transcript Translating Translation Initiation Translations cell type developmental plasticity healing induced pluripotent stem cell insight male multicatalytic endopeptidase complex mutant neuronal cell body novel pluripotency prevent programs protein degradation protein expression scaffold sperm cell synergism therapeutic target tissue regeneration tumor tumorigenesis

项目摘要

Project Summary Germ cells and somatic cells from an individual carry identical copies of DNA; yet, only germ cells have the potential to give rise to all the cell types of each subsequent generation. This suggests that epigenetic factors confer pluripotent and immortal potential to germ cells. Some of these epigenetic factors are exclusive to the germ cell cytoplasm, and in certain cases their presence is enough to reprogram somatic nuclei to restore pluripotency and immortal potential. A distinguishing feature of germ cell cytoplasm are germ granules. Germ granules are a phase-separated, heterogeneous mix of RNA and protein that have been observed in the germline of most animals. Because core germ-granule composition is conserved from nematodes to humans, the genetically tractable model C. elegans is used to study how germ granules function to regulate the pluripotent and immortal potential of germ cells. Germ granules extend the environment of the nuclear pore into the cytoplasm, where they serve as a safety next to ensure that nascent transcripts coming from the nucleus are licensed for germline expression. Depletion of germ granules in C. elegans causes sterility and germ-to-soma transformation. The long-term objectives of this research are to understand how germ granules regulate cellular pluripotency and their potential to be manipulated to induce pluripotency when it is needed (e.g. regeneration) or shut it off when it is not (e.g. cancer). A conserved core of germ-granule proteins act as multipotency factors, and are often repurposed in the soma during development, tissue regeneration, and tumorigenesis. This core consists of Vasa DEAD-box RNA helicases, small RNA binding Argonaute proteins, and LOTUS-Tudor domain proteins that interface and stimulate Vasa and Argonaute activity. The role of these multipotency factors during tumorigenesis, and whether they contribute to the phenomenon of cancer/testis (CT) antigen expression in various tumors has not been explored. Here, C. elegans is used to understand how these core proteins are expressed and distributed, what they interact with, and their role in fertility and developmental plasticity. The specific aims in this proposal look at 1) the role of germ granules in protein expression and turnover, primarily focusing on the novel interaction of GLH/Vasa with PCI scaffolding complexes, 2) how GLH promotes the differential translation of spermatogenic transcripts, and 3) how GLH and a new LOTUS-Tudor protein called LOTR-1 cooperates to maintain germline integrity. If successful, these findings will reveal novel ways to manipulate cellular pluripotency and expose the potential therapeutic targets that directly regulate the cytoplasm instead of nuclear gene expression networks.

项目概要个体的生殖细胞和体细胞携带相同的 DNA 拷贝；然而，只有生殖细胞具有产生后续每一代所有细胞类型的潜力。这表明表观遗传因素赋予生殖细胞多能和永生的潜力。其中一些表观遗传因素是生殖细胞细胞质，在某些情况下，它们的存在足以重新编程体细胞核以恢复多能性和不朽的潜力。生殖细胞细胞质的一个显着特征是生殖颗粒。胚芽颗粒是在种系中观察到的 RNA 和蛋白质的相分离、异质混合物大多数动物。由于核心胚芽颗粒成分从线虫到人类都是保守的，遗传易处理模型秀丽隐杆线虫用于研究胚芽颗粒如何发挥调节多能性的作用以及生殖细胞永生的潜力。胚芽颗粒将核孔的环境扩展到细胞质，它们作为安全的旁边，以确保来自细胞核的新生转录本获得生殖系表达许可。秀丽隐杆线虫中胚芽颗粒的消耗导致不育和胚芽转化为体转变。这项研究的长期目标是了解细菌颗粒如何调节细胞多能性及其在需要时（例如再生）被操纵以诱导多能性的潜力或者在不正常时将其关闭（例如癌症）。胚芽颗粒蛋白的保守核心充当多能因子，并且经常在发育、组织再生和肿瘤发生过程中的体细胞。该核心由 Vasa DEAD-box RNA 组成解旋酶、小 RNA 结合 Argonaute 蛋白和 LOTUS-Tudor 结构域蛋白刺激瓦萨号和阿尔戈号的活动。这些多能因子在肿瘤发生过程中的作用，以及是否它们导致各种肿瘤中癌症/睾丸（CT）抗原表达的现象尚未被证实探索过。在这里，利用秀丽隐杆线虫来了解这些核心蛋白是如何表达和分布的，什么它们与生育力和发育可塑性相互作用并发挥作用。该提案的具体目标如下 1）胚芽颗粒在蛋白质表达和周转中的作用，主要关注 GLH/Vasa 与 PCI 支架复合物，2) GLH 如何促进生精的差异翻译转录本，以及 3) GLH 和一种名为 LOTR-1 的新 LOTUS-Tudor 蛋白如何合作维持种系正直。如果成功，这些发现将揭示操纵细胞多能性的新方法并揭示直接调节细胞质而不是核基因表达网络的潜在治疗靶点。

项目成果

期刊论文数量（4）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

acdh-1 transcripts are expressed in the intestine.

acdh-1 转录物在肠道中表达。

DOI：
10.17912/w2v365
发表时间：
2017
期刊：
microPublication biology
影响因子：
0
作者：
Updike,Dustin
通讯作者：
Updike,Dustin

Quelling germ cell pluripotency on the genital ridge.

抑制生殖嵴上的生殖细胞多能性。

DOI：
10.1073/pnas.1918899116
发表时间：
2019
期刊：
Proceedings of the National Academy of Sciences of the United States of America
影响因子：
11.1
作者：
Updike,DustinL
通讯作者：
Updike,DustinL

swt-3 transcripts are expressed in Z1/Z4 during embryogenesis and in the adult spermatheca.

swt-3 转录本在胚胎发生期间和成体受精囊中的 Z1/Z4 中表达。