Defining Activities of KDMS Essential to Development and Viability

摘要 赖氨酸去甲基酶5(KDM5)家族的转录调控对正常发育很重要， 而它们的失调是导致智力残疾和多种癌症的关键因素。大多数工作都是最新的 专注于KDM5蛋白的组蛋白去甲基酶活性，其目标是活性染色质标记 H3K4me3。然而，KDM5蛋白也可以通过非酶机制调节转录。而当 KDM5对发育是必不可少的，其去甲基酶活性不是必需的，正如生存能力所证明的那样 去甲基酶--死亡的成年苍蝇。在这项工作中，我们将通过一种新的截断来研究KDM5的基本功能 等位基因kdm5Q19，不改变脱甲基酶活性。Kdm5Q19在先前的 在KDM5的C末端的一个内在无序区域内，未被识别的进化保守的基序。 Kdm5q19动物不能存活到成年，这与Null、去甲基酶死亡和其他突变体不同 在我们的实验室中产生的，表明被截断破坏的基序具有一个重要的作用尚不清楚 在正常的KDM5函数中。为了进一步剖析KDM5这个区域的分子活性，我们生成了 Kdm5的额外等位基因，以提炼蛋白质的关键区域(S)，并评估其活性和变化 抄写。 在这项工作中，我们将(1)定义KDM5内生存所需的基本区域，并描述它们的作用 在发展中(2)定义由KDM5的C末端赋予的基本分子功能。一起， 这些研究将提炼KDM5C末端的临界区(S)，并确定这些关键区域的正常作用 转录调控区域。

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.