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.

DNA 由组蛋白包装到染色质中，以适应细胞核。可以分为开放的、转录活跃的区室和封闭的、转录沉默的区室。不同的非组蛋白染色质结合蛋白有助于整体染色质重塑。酶、转录因子和转录相关 RNA 对于解凝很重要基因组的转录活性部分和相关的非编码RNA。对于基因组转录非活性部分的浓缩非常重要。组织和压实对于发育和染色体过程中细胞状态的变化至关重要有丝分裂期间的分离。在有丝分裂开始时，染色体结构发生巨大变化，因为染色体紧密且当细胞进入有丝分裂时，细胞中存在的所有结构都进行个体化。间期核结构的擦除与逐步去除相关。粘连蛋白复合物的形成和个体化是通过组合完成的。凝缩蛋白复合物和拓扑异构酶 IIα 的作用。结构在时间上与有丝分裂染色体末端的转录抑制相关。簇和去凝聚以重组单个染色体间核。通过在有丝分裂过程中从染色质中去除的许多不同蛋白质的重新结合来完成与核转录的恢复在时间上相关。有丝分裂期间染色质结构的变化与核的变化相关转录活性，但我们最近发现这两个过程之间的联系却知之甚少。前期去除染色体中的粘连蛋白复合物对于沉默有丝分裂转录至关重要有丝分裂过程中染色质结合 RNA 从染色体上的去除是由磷酸化介导的 SAF-A 的鉴定将染色体结构的变化与染色体的变化联系起来。转录活动提供了一个了解这些事件如何相互关联的机会。将重建由 SAF-A 介导的核酸相互作用的生物化学，以提供关于如何然后我们将研究如何从这种丰富的蛋白质中去除 SAF-A。然后我们将研究 SAF- 的重新结合。有丝分裂末期的染色质 A 促进核重组和转录激活。本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分离和转录调控。