Characterizing the LINE-1 Retrotransposition-Replication Conflict

表征 LINE-1 逆转录转座-复制冲突

基本信息

批准号：
10634604
负责人：
KATHLEEN H BURNS
金额：
$ 39.68万
依托单位：
DANA-FARBER CANCER INST
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-07-15 至 2025-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10634604
关键词：
Affect BARD1 gene BRCA1 gene Binding Biochemical Biological Assay CRISPR/Cas technology Cancer Cell Growth Cell Nucleus Cell Survival Cell physiology Cells Code Complement Complex Conflict (Psychology)Cytostatics DNA DNA Repair DNA Repair Gene DNA Repair Pathway DNA Transposable Elements DNA biosynthesis DNA replication fork Data Detection Fanconi Anemia-BRCA Pathway Fanconi&apos s Anemia Fiber Foundations Funding Opportunities Gene Mutation Genes Genome Genomics Human Immunofluorescence Immunologic Inhibition of Cancer Cell Growth International Knock-out Length Lethal Genes Ligation Long Interspersed Elements Malignant Neoplasms Maps Mediating Mediator Methods Methylation Mobile Genetic Elements Molecular Mutate Mutation Normal Cell Nuclear Open Reading Frames Pathway interactions Poly(ADP-ribose) Polymerase Inhibitor Poly(ADP-ribose) Polymerases Process Proteins RNA-Binding Proteins RNA-Directed DNA Polymerase Research Retrotransposition Retrotransposon Reverse Transcription Role S phase Series Short Interspersed Nucleotide Elements Signal Pathway Signal Transduction Site System Testing The Cancer Genome Atlas Therapeutic Tumor Suppressor Genes Visualization biological adaptation to stress cancer cell cancer genome cell growth coping cytotoxic endonuclease experimental study fitness genome-wide in vivo mutant next generation sequencing overexpression prevent promoter replication stress restraint synthetic lethal interaction translocase ubiquitin ligase

项目摘要

Project Summary Much of our genome is made up of interspersed repeats derived from the activities of mobile genetic elements. There are subsets of these sequences that become activated in cancers. Our research lab and others have shown that nearly half of cancers fail to restrain long interspersed element-1 (LINE-1, L1) sequences. L1 is an active retrotransposon that codes for two proteins, an RNA binding protein (ORF1p) and a protein with endonuclease and reverse transcriptase activities (ORF2p). Paradoxically, though these proteins are pervasively expressed in many human cancers, they inhibit cell growth in culture. We have exciting new data from genome-wide knockout screens demonstrating how tumor suppressor gene mutations found in cancers enable them to survive and grow despite their expression of L1. More importantly, these screens have also identified genes that become essential in cells coping with L1 expression. These synthetic lethal genes represent unique molecular vulnerabilities for L1(+) cells. L1(+) cells require specific DNA repair pathways, effective replication stress signaling pathways, and replication fork restart capabilities. Here, we will test the hypothesis that these pathways are required to eliminate L1 insertion intermediates and prevent collisions between these intermediates and DNA replication forks. This project will increase our understanding of how cancer cells replicate their DNA. It could lay a foundation to leverage L1-associated DNA replication stress to limit cancer cell growth.

项目概要我们的基因组大部分是由源自移动遗传元件活动的散布重复序列组成。这些序列的某些子集在癌症中被激活。我们的研究实验室和其他实验室有研究表明，近一半的癌症无法抑制长散布元件 1 (LINE-1，L1) 序列。 L1 是一个活性逆转录转座子编码两种蛋白质，一种是 RNA 结合蛋白 (ORF1p)，另一种是具有核酸内切酶和逆转录酶活性（ORF2p）。矛盾的是，尽管这些蛋白质它们在许多人类癌症中普遍表达，可抑制培养物中的细胞生长。我们有令人兴奋的新数据来自全基因组敲除筛选，展示了如何在癌症中发现肿瘤抑制基因突变使他们能够生存和成长，尽管他们表达 L1。更重要的是，这些屏幕还鉴定出在处理 L1 表达的细胞中至关重要的基因。这些合成的致死基因代表 L1(+) 细胞独特的分子脆弱性。 L1(+)细胞需要特定的DNA修复途径，有效的复制压力信号通路和复制叉重启功能。在这里，我们将测试假设需要这些途径来消除 L1 插入中间体并防止碰撞这些中间体和 DNA 复制叉之间。这个项目将加深我们对如何癌细胞复制它们的DNA。它可以为利用 L1 相关的 DNA 复制压力奠定基础限制癌细胞生长。