Analysis of chromatin-RNA interactions during the cell cycle.

细胞周期中染色质-RNA 相互作用的分析。

基本信息

批准号：
10338329
负责人：
Michael Demian Blower
金额：
$ 33万
依托单位：
BOSTON UNIVERSITY MEDICAL CAMPUS
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-03-16 至 2025-12-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10338329
关键词：
ATPase Domain Affect Architecture Binding Binding Proteins Biochemical Biological Cell Cycle Cell Line Cell Nucleus Cell division Cells Chromatin Chromatin Loop Chromatin Structure Chromosome Condensation Chromosome Segregation Chromosome Structures Chromosomes Complex DNA DNA Polymerase II Development Enzymes Euchromatin Event Excision Gene Expression Gene Expression Regulation Genetic Genetic Epistasis Genetic Transcription Genome Genome Stability Genomic approach Genomics Goals Heterochromatin Histones Human Human Development Image In Vitro Interphase Interphase Chromosome Knowledge Link Malignant Neoplasms Mediating Messenger RNA Mitosis Mitotic Mitotic Chromosome Molecular Mus Mutation Neurodevelopmental Disorder Nuclear Nuclear Protein Nuclear Structure Nucleic Acid Biochemistry Nucleic Acids Pathway Analysis Pathway interactions Phenotype Phosphorylation Physical condensation Preparation Process Prophase Proteins RNA RNA Binding RNA Splicing Regulation Sister Chromatid Structure Surface Time Transcriptional Activation Transcriptional Regulation Untranslated RNA Work cancer cell chromatin remodeling cohesin condensin de novo mutation design experimental study genetic analysis genetic information genetically modified cells human error insight non-histone protein programs reconstitution segregation telophase transcription factor

项目摘要

DNA is packaged by histone proteins into chromatin in order to fit into the nucleus of a cell. Globally, chromatin can be separated into open, transcriptionally active and closed, transcriptionally silent compartments. Many different nonhistone, chromatin-binding proteins contribute to global chromatin structure. Chromatin remodeling enzymes, transcription factors, and transcription-associated RNAs are important for decondensing transcriptionally active portions of the genome. Heterochromatin proteins and associated noncoding RNAs are important for condensation of the transcriptionally inactive portions of the genome. Changes in chromatin organization and compaction are critical for cell state changes during development and chromosome segregation during mitosis. Chromosome structure changes dramatically at the beginning of mitosis as chromosomes compact and individualize in preparation for segregation. As cells enter into mitosis all of the structure present in the interphase nucleus is erased. Erasure of interphase nuclear structure is correlated with the stepwise removal of the Cohesin complex. Chromosome condensation and individualization are accomplished by the combined action of the Condensin complexes and Topoisomeriase IIα. Interestingly, dramatic changes in chromosome structure are temporally correlated with suppression of transcription. At the end of mitosis chromosomes cluster and decondense to reform a single interphase nucleus. Chromosome decondensation and clustering is accomplished by the rebinding of many different proteins that are removed from chromatin during mitosis and is temporally correlated with the resumption of nuclear transcription. While changes in chromatin structure during mitosis are correlated with changes in nuclear transcriptional activity, little is known about how the two processes are linked. We have recently discovered that prophase removal of the Cohesin complex form chromosomes is critical for silencing mitotic transcription and that removal of chromatin-bound RNAs from chromosomes during mitosis is mediated by phosphorylation of SAF-A. The identification of molecular pathways linking changes in chromosome structure to changes in transcriptional activity presents an opportunity to understand how these events are linked. In this proposal we will reconstitute the biochemistry of nucleic acid interactions mediated by SAF-A to provide a picture of how this abundant protein controls chromosome structure. We will then examine how removal of SAF-A from chromatin promotes chromosome condensation during prophase. We will then examine how rebinding of SAF- A to chromatin at the end of mitosis promotes nuclear reformation and transcriptional activation. Collectively, the experiments outlined in this proposal will provide new insight into changes in chromatin-RNA interactions that control chromatin structure and how these changes contribute to accurate chromosome segregation and transcriptional regulation.

通过组蛋白将DNA包装到染色质中，以适合细胞的核。全球，染色质可以分为开放，转录活跃和封闭的，转录的无声隔室。许多不同的非组蛋白，染色质结合蛋白有助于整体染色质结构。染色质重塑酶，转录因子和转录相关的RNA对于反应很重要基因组的转录活性部分。异染色质蛋白和相关的非编码RNA是对于基因组的转录不活动部分的凝结很重要。染色质的变化组织和压实对于发育和染色体过程中的细胞状态变化至关重要有丝分裂过程中的分离。染色体结构在有丝分裂开始时随着染色体紧凑而发生巨大变化个性化准备隔离。当细胞进入有丝分裂时，所有存在的结构相间核us被删除。擦除相间核结构与逐步删除相关粘蛋白复合物的。染色体凝结和个性化是通过合并的冷凝蛋白复合物和拓扑异构体IIα的作用。有趣的是，染色体的戏剧性变化结构与转录的抑制暂时相关。有丝分裂染色体的结尾染色体反应和聚类是通过在有丝分裂和与核转录的恢复暂时相关。虽然有丝分裂过程中染色质结构的变化与核的变化相关转录活动，关于这两个过程的链接知之甚少。我们最近发现了预言去除粘蛋白复合物形式染色体对于沉默有丝分裂转录至关重要并且在有丝分裂过程中从染色体中去除染色质的RNA是由磷酸化介导的 Saf-a。鉴定分子途径将染色体结构变化与变化联系起来的变化转录活动为了解这些事件的联系方式提供了机会。在这个建议中，我们将重建由SAF-A介导的核酸相互作用的生物化学，以提供有关如何这种丰富的蛋白质控制染色体结构。然后，我们将研究如何从SAF-A中删除染色质促进预言期间染色体凝结。然后，我们将研究SAF的重新启动有丝分裂结束时的染色质促进核识别和转录激活。共同该提案中概述的实验将为染色质-RNA相互作用的变化提供新的见解控制染色质结构以及这些变化如何有助于准确的染色体隔离和转录调节。