FUNCTIONAL CHARACTERIZATION OF THE PRDM10-ZN FINGER TRANSCRIPTION FACTOR IN EARLY MAMMALIAN DEVELOPMENT

PRDM10-ZN 指转录因子在早期哺乳动物发育中的功能特征

• 批准号: 10477941
• 负责人: Ernesto Guccione
• 金额: $ 36.34万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-31 至 2026-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10477941
• 关键词: AT Rich Sequence; Address; Adult; Aging; Apoptosis; Automobile Driving; Binding; Biological Models; Catalytic Domain; Cell Cycle Progression; Cell physiology; Cells; Chromatin; Chromatin Structure; Communication; Complex; Couples; Cues; DNA; DNA Binding; DNA Binding Domain; Data; Defect; Developed Countries; Development; Developmental Process; Diagnosis; Disease; Drops; Embryo; Embryonic Development; Event; Family; Family Planning; Family member; Fertility; Fingers; Future; Gene Activation; Gene Expression; Genes; Genetic; Genetic Transcription; Germ Cells; Glutamine; Herpes zoster disease; Infertility; Knowledge; Link; Lysine; Malignant Fibrous Histiocytoma; Methyltransferase; Mitotic; Modeling; Mus; Nature; Oncogenic; Oocytes; Oogenesis; Pathologic; Pathway interactions; Patients; Pharmacology; Phenotype; Pre-implantation Embryo Development; Pregnancy Complications; Process; Property; Proteins; Regulation; Reproductive Biology; Reproductive Medicine; Research; Role; SET Domain; Science; Signal Transduction; Societies; Soft tissue sarcoma; Time; Tissues; Transcription Coactivator; Transcriptional Activation; Transcriptional Regulation; Translational Regulation; Translations; Vertebrates; Work; base; embryonic stem cell; epigenetic regulation; experimental study; extracellular; gene network; idiopathic infertility; inhibitor; member; paralogous gene; programs; promoter; protein expression; side effect; stem cell biology; transcription factor; zygote

SUMMARY During embryonic development and lineage specification, a complex set of extracellular signals is integrated on chromatin by Master Transcription Factors (TF) that modify chromatin structure and gene expression. PRDM proteins are TFs, containing a PR-methyltransferase domain at their N-terminus and DNA-binding Zn-fingers at their C-terminus. All 17 PRDM family members are well known for their role as master regulator of lineage specification and their deregulated expression is often linked to diseases. Our group has characterized the function of PRDM10, a transcriptional activator, orchestrating mouse preimplantation development. Our preliminary data indicate that: 1) Prdm10 zygotic KO embryos have visible defects at embryonic day E3.5; 2) Prdm10 maternal (Zona Pellucida3 (Zp3)-CRE) KO embryos arrest at the zygote/two-cell stage; 3) PRDM10 is unique among PRDM family members, as it contains a Glutamine-rich coactivator domain at its C-terminus. Based on this preliminary data we propose the following experiments: In Aim1 we will determine the function of maternal PRDM10. We will take advantage of the Zp3-CRE strain, to delete Prdm10 in the oocyte, and deplete the PRDM10 maternal protein contribution in the zygote. This will allow us to study the function of PRDM10 before zygotic transcriptional activation and determine the gene network directly regulated by PRDM10. In Aim2 we will determine the mechanism of action of PRDM10 as a transcription factor. In this aim we will characterize the mode of action of PRDM10 (i.e. its methyltransferase activity, its protein interactome, and its transcriptional activation and DNA binding-properties). We will also specifically address PRDM10’s function in dictating the choice of alternative promoter usage. Finally, we will validate the function in oocyte and early embryogenesis of key PRDM10-downstream effectors. The significance of these studies is that Prdm10 is an uncharacterized maternal effect gene, and knowledge of the pathways regulated by this Zn-finger TF will be useful to the field of stem cell biology and reproductive medicine.

概括 在胚胎发育和谱系规范期间，整合了一组复杂的细胞外信号 通过主转录因子（TF）修饰染色质结构和基因表达的染色质。 prdm 蛋白质是TFS，在其N末端和DNA结合Zn-Finger处包含PR-甲基转移酶结构域 他们的C末端。所有17名PRDM家庭成员都以其作为血统总监管者的角色而闻名 规范及其放松管制的表达通常与疾病有关。 我们的小组已经表征了prdm10的功能，prdm10是一个转录激活器，编排鼠标的功能 植入前的发展。我们的初步数据表明：1）PRDM10合子KO胚胎可见 胚胎日E3.5的缺陷； 2）prdm10物质（Zona Pelucida3（ZP3）-CRE）KO胚胎在The The The The The The Zygote/Twiper阶段； 3）PRDM10在PRDM家庭成员中是独一无二的，因为它含有谷氨酰胺丰富 共激活因子域在其C末端。基于此初步数据，我们提出了以下实验： 在AIM1中，我们将确定母体PRDM10的功能。我们将利用zp3-cre 应变，以删除卵母细胞中的PRDM10，并取代合子中的PRDM10母体蛋白质贡献。这 将允许我们在合子转录激活之前研究PRDM10的功能并确定基因 网络直接由PRDM10调节。 在AIM2中，我们将确定PRDM10作为转录因子的作用机理。在这个目标中我们 将表征PRDM10的作用方式（即其甲基转移酶活性，其蛋白质相互作用组和 其转录激活和DNA结合概念）。我们还将专门解决PRDM10的功能 在决定选择替代启动子使用情况时。最后，我们将验证卵母细胞和早期的功能 关键PRDM10-下游效应的胚胎发生。 这些研究的意义在于，PRDM10是一个未表征的遗传效应基因，知识是 该Zn手指TF调节的途径将对干细胞生物学和生殖有用 药品。