The Role of LEM Domain Proteins in Nuclear Function

LEM 结构域蛋白在核功能中的作用

基本信息

批准号：
10175883
负责人：
PAMELA K. GEYER
金额：
$ 14.81万
依托单位：
UNIVERSITY OF IOWA
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-06-01 至 2023-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10175883
关键词：
ATR checkpoint Adult Affect Binding Binding Proteins CHEK2 gene Cell Death Cell Maintenance Cell Survival Cells Cessation of life Chromatin Clustered Regularly Interspaced Short Palindromic Repeats Complex Cyst Cytology DNA DNA Damage Data Defect Deformity Development Disease Disease model Drosophila genus Failure Family Fatty acid glycerol esters Functional disorder Genes Genetic Genetic Transcription Germ Cells Health Heterochromatin High-Throughput RNA Sequencing Homeostasis Human Lead Link Membrane Monitor Mutation Nuclear Nuclear Inner Membrane Nuclear Lamina Nuclear Structure Ontology Pathway interactions Phenotype Phosphotransferases Play Premature aging syndrome Proteins Role Shapes Signal Pathway Signal Transduction Skeletal Muscle Skin Stress Structural defect Structure Tertiary Protein Structure Testing Tissues Transcriptional Activation Variant adult stem cell barrier-to-autointegration factor base bone delta protein ds-DNA experimental study germline stem cells histone-binding proteins improved knock-down mutant novel overexpression prevent response sensor stem cell homeostasis stem cell population stem cells therapeutic development tool transcription factor transcriptome

项目摘要

Project Summary Human laminopathies are caused by mutations in genes encoding nuclear lamina (NL) proteins. A unifying disease model suggests that lost tissue homeostasis is due to a failure to maintain adult stem cells. Although NL proteins responsible for laminopathies have been identified, it remains unclear how these proteins maintain healthy stem cell populations and promote tissue homeostasis. The conserved NL family of LEM-domain (LEM-D) proteins play a critical role in building nuclear structure and the NL. LEM-D proteins bind Barrier-to-Autointegration Factor (BAF), a double stranded DNA and histone binding protein. We investigate the Drosophila LEM-D family, focusing on Otefin, a LEM-D protein that is required for survival of adult germline stem cells (GSCs). The otefin mutant GSCs carry structural deformities of the NL and chromatin changes that are shared with laminopathic cells. My lab discovered that these mutant GSCs die because of activation of a novel checkpoint pathway that uses two DNA damage response (DDR) kinases, ATR and Checkpoint kinase 2 (Chk2). Although otefin mutant GSCs carry DNA damage, damage accumulation depends upon Chk2, demonstrating that DNA damage results from checkpoint activation. Based on these and other data, we hypothesize that NL deformation is responsible for activation of ATR and Chk2, a prediction supported by emerging evidence that ATR is a global sensor of structural deformities of cellular components. In this proposal, two Aims are proposed. In Aim 1, we will define the mechanism of ATR/Chk2 activation in otefin mutant GSCs. In Aim 2, we define Chk2-dependent pathways involved in GSC death. Nuclear shape changes are shared features of laminopathies and premature aging syndromes. We predict that activation of the NL checkpoint might contribute to lost stem cell maintenance in these diseases.

项目概要人类核纤层蛋白病是由编码核纤层 (NL) 蛋白的基因突变引起的。统一的疾病模型表明，组织稳态的丧失是由于未能维持成体干细胞。尽管已鉴定出导致核纤层蛋白病的 NL 蛋白，但它目前尚不清楚这些蛋白质如何维持健康的干细胞群并促进组织稳态。 LEM 结构域 (LEM-D) 蛋白的保守 NL 家族在在构建核结构和 NL 中的作用。 LEM-D 蛋白结合自动整合屏障因子 (BAF)，一种双链 DNA 和组蛋白结合蛋白。我们研究果蝇 LEM-D 家族，专注于 Otefin，一种成年种系存活所需的 LEM-D 蛋白干细胞（GSC）。 otefin 突变体 GSC 携带 NL 和染色质的结构畸形与核纤层蛋白病细胞共有的变化。我的实验室发现这些突变的 GSC 会死亡因为使用两种 DNA 损伤反应的新型检查点途径被激活 (DDR) 激酶、ATR 和检查点激酶 2 (Chk2)。尽管otefin突变体GSC携带DNA 损伤，损伤累积取决于Chk2，证明DNA损伤导致从检查点激活。基于这些和其他数据，我们假设 NL 变形负责 ATR 和 Chk2 的激活，这一预测得到了新出现的证据的支持： ATR 是细胞成分结构变形的全局传感器。在这个提案中，两个提出了目标。在目标 1 中，我们将定义 otefin 中 ATR/Chk2 激活的机制突变的 GSC。在目标 2 中，我们定义了参与 GSC 死亡的 Chk2 依赖性途径。核形状变化是核纤层病和早衰综合征的共同特征。我们预测 NL 检查点的激活可能导致干细胞维持的丧失这些疾病。