Post-translational regulation of sperm development and function in C. elegans

秀丽隐杆线虫精子发育和功能的翻译后调控

• 批准号: 10653491
• 负责人: DIANE CAROL SHAKES
• 金额: $ 41.22万
• 依托单位: COLLEGE OF WILLIAM AND MARY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-04-01 至 2026-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10653491
• 关键词: Affect; Alzheimer' s Disease; Alzheimer' s disease pathology; Biochemical; Biological Models; Biology; Bypass; CRISPR/Cas technology; CSNK1A1 gene; Caenorhabditis elegans; Cell Differentiation process; Cell physiology; Cells; Cellular biology; Chromosomes; Collaborations; Complement; Cytoskeletal Proteins; Defect; Development; Developmental Biology; Drug Targeting; Environment; Enzymes; Escherichia coli; Female; Fertility; Fertilization; Gene Duplication; Gene Family; Gene Fusion; Genes; Genetic; Genetic Screening; Genetic Transcription; Giant Cells; Goals; Homologous Gene; Human; Immunofluorescence Immunologic; Impairment; In Vitro; Individual; Infertility; Knock-out; Malignant Neoplasms; Meiosis; Mentors; Metaphase; Microscope; Molecular; Monitor; Morphology; Mutation; Nematoda; Oogenesis; Outcome; Parasites; Pathogenicity; Pathologic; Pathway interactions; Pattern; Persons; Pharmacologic Substance; Phenotype; Phosphoric Monoester Hydrolases; Phosphotransferases; Post-Translational Regulation; Process; Proteins; Publishing; Regulation; Research; Residual state; Role; Science; Site; Sperm Motility; Spermatocytes; Spermatogenesis; Students; System; Testing; Training; Transgenic Organisms; Uterus; Work; casein kinase I; cell type; experimental study; fundamental research; gene conservation; human disease; imaging system; in vivo; insight; knock-down; knockout gene; male; member; migration; mutant; novel; programs; protein function; reproductive tract; segregation; sperm cell; sperm function; sperm protein; tau-tubulin kinase; tool; training opportunity; undergraduate research experience; undergraduate student; ward

RELEVANCE: Under both normal and pathological conditions, kinases and phosphatases are key post-translational regulators of cellular and developmental pathways. Mis-regulation of these enzymes is associate with diverse pathologies from Alzheimer's and cancer to infertility. Our studies will investigate a kinase SPE-6 that is related to the Alzheimer's relevant kinase (tau tubulin kinase and a pseudophosphatase SPE-54, a class of enzymes whose critical roles are only recently being recognized. RATIONALE: Our studies will investigate the function of these proteins in the unique context of C. elegans sperm development and sperm function. Because humans and C. elegans share many of the same genes, discoveries made in C. elegans can be directly relevant to humans. Conversely, nematode-specific proteins required for fertility are promising drug targets for sterilizing nematode parasites that infect over a billion people. As an experimental system, the molecular and genetic tools of C. elegans including the ability to monitor sperm migration in vivo through its transparent body, make it an outstanding model system for fundamental research. Furthermore, the unique biology of its spermatogenesis program provides a novel context for exploring the function of kinases and phosphatases, largely in the absence of new gene transcription. OBJECTIVES: The proposed studies will explore the functions of a kinase SPE-6 and a pseudophosphatase SPE-54 whose phenotypic defects provide an entry point for investigating largely unexplored cellular and developmental pathways as such analysis often generates transformative insights in biology. First, we will investigate the molecular mechanisms by which SPE-6 regulates three distinct cellular pathways: a) assembly of a nematode specific cytoskeletal protein into tightly packed fibrous bodies, b) detachment of spermatocytes from the germline syncytium, and c) the remodeling of chromosomes so that they can be segregated during the meiotic divisions. Secondly, we will use both in vivo and in vitro approaches to investigate how SPE-54 regulates the directed mobility of the sperm to the site of fertilization. The findings should broadly inform our understanding of kinase and pseudophosphatase functions in diverse cell types.

相关性：在正常和病理条件下，激酶和磷酸酶是 细胞和发育途径的关键翻译后调节因子。监管不善 这些酶与从阿尔茨海默氏症、癌症到不孕不育的各种病理因素有关。 我们的研究将研究与阿尔茨海默氏症相关的激酶(Tau)相关的一种激酶SPE-6 微管蛋白激酶和伪磷酸酶SPE-54，这类酶的关键作用是 直到最近才被认出。理论基础：我们的研究将调查这些基因的功能 蛋白质在线虫精子发育和精子功能的独特背景下。因为 人类和线虫有许多相同的基因，在线虫中发现的可能是 与人类直接相关。相反，繁殖力所需的线虫特异性蛋白质是 有希望的药物目标是消毒感染超过10亿人的线虫寄生虫。作为一种 实验系统，线虫的分子和遗传工具，包括 通过透明的身体监测精子在体内的迁移，使其成为杰出的典范 基础研究体系。此外，其精子发生的独特生物学 程序为探索激酶和磷酸酶的功能提供了新的背景， 很大程度上是在没有新的基因转录的情况下。目标：拟议的研究将 表型为SPE-6和SPE-54的激酶SPE-6和伪磷酸酶SPE-54的功能研究 缺陷为研究大部分未被探索的细胞和发育提供了一个切入点 像这样的分析途径通常会产生生物学上的变革性洞察。首先，我们将 探讨SPE-6调控三种不同细胞的分子机制 途径：a)将线虫特异性细胞骨架蛋白组装成紧密堆积的纤维 B)精母细胞从生殖系合体中分离，以及c)精母细胞的重塑。 染色体，以便它们可以在减数分裂过程中分离。其次，我们将 使用体内和体外方法研究SPE-54是如何调节定向的 精子到受精部位的移动性。这些发现应该会广泛地告诉我们 了解不同细胞类型中的激酶和伪磷酸酶的功能。