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.

总结 在胚胎发育和谱系特化过程中，一组复杂的细胞外信号被整合 主转录因子（TF）修饰染色质结构和基因表达。PRDM 蛋白质是TF，在其N末端含有PR-甲基转移酶结构域，在其N末端含有DNA结合锌指， 他们的C末端PRDM家族的所有17名成员都因其作为血统主要调节者的角色而闻名 基因特化及其失调表达通常与疾病有关。 我们的小组已经表征了PRDM 10的功能，PRDM 10是一种转录激活因子， 着床前发育我们的初步数据表明：1）Prdm 10合子KO胚胎具有可见的 2）Prdm 10母体（透明带3（Zp 3）-CRE）KO胚胎在E3.5天停滞， 3）PRDM 10在PRDM家族成员中是独特的，因为它含有富含谷氨酰胺的蛋白质。 在其C-末端的辅激活因子结构域。基于这些初步数据，我们提出以下实验： 在Aim 1中，我们将确定母体PRDM 10的功能。我们将利用Zp 3-CRE 菌株，以删除卵母细胞中的Prdm 10，并耗尽受精卵中的PRDM 10母体蛋白贡献。这 这将使我们能够研究PRDM 10在合子转录激活前的功能，并确定基因 由PRDM 10直接管理的网络。 在Aim 2中，我们将确定PRDM 10作为转录因子的作用机制。为此，我们 将表征PRDM 10的作用模式（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结合性质）。我们还将专门讨论PRDM 10的功能 在决定替代启动子使用的选择方面。最后，我们将在卵母细胞和早期验证其功能。 关键PRDM 10下游效应子的胚胎发生。 这些研究的意义在于Prdm 10是一种未表征的母体效应基因，并且知识 这种锌指TF调控的途径将有助于干细胞生物学和生殖领域 药