Defining Activities of KDMS Essential to Development and Viability

定义对发展和生存至关重要的 KDMS 活动

基本信息

批准号：
10672661
负责人：
Melissa Castiglione
金额：
$ 4.77万
依托单位：
ALBERT EINSTEIN COLLEGE OF MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-03-30 至 2026-03-29
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10672661
关键词：
Address Adult Alleles Amino Acid Sequence Amino Acids Animals Binding Biological Models C-terminal Characteristics Chromatin Chromatin Remodeling Factor Conserved Sequence Control Animal Data Defect Development Developmental Delay Disorders Disease Drosophila genus Family Gene Expression Gene Expression Regulation Genes Genetic Transcription Genomics Gland Goals Histones Homologous Gene Human Intellectual functioning disability Knowledge Label Lysine Malignant Neoplasms Mediating Methylation Molecular Neurosecretory Systems Nonsense Mutation Pathway interactions Phenotype Play Point Mutation Protein Family Protein Region Proteins Proteomics Publishing Regulation Regulator Genes Role Structure Study models Terminator Codon Testing Tissues Work alpha helix design feature detection fly histone demethylase in vivo molecular recognition mutant neuron development novel paralogous gene programs protein protein interaction recruit targeted treatment tool transcriptome sequencing transcriptomics

项目摘要

ABSTRACT The Lysine demethylase 5 (KDM5) family of transcriptional regulators are important for normal development, and their dysregulation is a key driver of intellectual disability and several forms of cancer. Most work to-date has focused on the histone demethylase activity of KDM5 proteins, which targets the active chromatin mark H3K4me3. However, KDM5 proteins can also regulate transcription through non-enzymatic mechanisms. While KDM5 is essential for development, its demethylase activity is not required, as is demonstrated by the viability of demethylase-dead adult flies. In this work, we will examine essential functions of KDM5 via a novel truncation allele, kdm5Q19, which does not alter demethylase activity. kdm5Q19 inserts a stop codon in a previously unrecognized, evolutionarily conserved, motif within an intrinsically disordered region of KDM5 at the C-terminus. kdm5Q19 animals do not survive to adulthood, which is distinct from null, demethylase dead, and other mutants generated in our lab, suggesting that the motif disrupted by the truncation has an essential as-yet-unknown role in normal KDM5 function. To further dissect the molecular activities of this region of KDM5, we generate additional alleles of kdm5 to refine the critical region(s) of the protein and assess viability and changes to transcription. In this work, we will (1) define essential regions within KDM5 required for viability, and to characterize their roles in development (2) define the essential molecular functions imparted by the C-terminus of KDM5. Together, these studies will refine the critical region(s) of the C-terminus of KDM5, and define the normal role of these regions in regulating transcription.

抽象的转录调节因子赖氨酸脱甲基酶 5 (KDM5) 家族对于正常发育非常重要，它们的失调是智力障碍和多种癌症的关键驱动因素。迄今为止大部分工作专注于 KDM5 蛋白的组蛋白去甲基化酶活性，该活性针对活性染色质标记 H3K4me3。然而，KDM5 蛋白也可以通过非酶机制调节转录。尽管 KDM5 对于发育至关重要，其去甲基酶活性不是必需的，正如活力所证明的那样去甲基酶死亡的成年果蝇。在这项工作中，我们将通过一种新颖的截断来检查 KDM5 的基本功能等位基因 kdm5Q19，不会改变去甲基酶活性。 kdm5Q19 在先前的密码子中插入一个终止密码子未识别的、进化上保守的基序，位于 KDM5 C 端本质上无序的区域内。 kdm5Q19 动物不能存活到成年，这与无效、去甲基酶死亡和其他突变体不同在我们的实验室中生成，表明被截断破坏的基序具有重要的但未知的作用在正常的 KDM5 功能中。为了进一步剖析 KDM5 该区域的分子活性，我们生成 kdm5 的其他等位基因，用于细化蛋白质的关键区域并评估活力和变化转录。在这项工作中，我们将 (1) 定义 KDM5 中生存所需的重要区域，并描述它们的角色开发中 (2) 定义 KDM5 C 末端赋予的基本分子功能。一起，这些研究将完善 KDM5 C 末端的关键区域，并确定这些区域的正常作用调控转录的区域。