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Mechanism and Function of Chromatin Positional Dynamics in Interphase

间期染色质位置动力学的机制和功能

基本信息

批准号：
8915221
负责人：
Alexandra Zidovska
金额：
$ 24.9万
依托单位：
NEW YORK UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-09-01 至 2017-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8915221
关键词：
ATP phosphohydrolase Acute leukemia Address Aftercare Antineoplastic Agents Award BRD2 gene Behavior Binding Biochemical Biochemistry Biological Biological Process Biology Bromodomain Cancer cell line Carcinoma Cell Cycle Regulation Cell Line Cell Nucleus Cell physiology Cells Cellular biology Centromere Chromatin Chromosome Territory Chromosomes Complement Complex Cultured Cells Cytoskeleton DNA DNA Packaging DNA Polymerase II DNA Repair DNA Topoisomerases Data Diagnostic Doctor of Philosophy Embryo Evolution Family Fellowship Foundations Future Gene Expression Genes Genetic Transcription Goals Hela Cells Histone H2B Histones Image In Situ Interphase Knowledge Label Lamins Laws Learning Life Malignant Neoplasms Mammalian Cell Maps Measurement Measures Mentors Methodology Methods Microscopy Modeling Molecular Molecular Biology Motor Multiple Myeloma Nature Nuclear Nuclear Fusion Nuclear Proteins Paper Pattern Phase Physics Poison Polymers Positioning Attribute Postdoctoral Fellow Property Protein Binding Proteins Proteomics Publications RNA Interference Research Resolution Role Site Spectrum Analysis System Testing Time Training Velocimetries Work abstracting analytical tool cancer cell cell motility cell type chromatin remodeling condensed matter physics fly in vivo inhibitor/antagonist knock-down particle particle spectroscopy physical model physical science post-doctoral training screening segregation single molecule skills small molecule telomere tool

项目摘要

Project Summary/Abstract The goal of the proposed research is (1) to measure spatially and temporally resolved chromatin dynamics in mammalian cells in interphase in vivo and (2) determine the origin and function of these dynamical behaviors by combining quantitative approaches derived from physical sciences with tools from molecular biology and biochemistry. The dynamic behavior of chromatin (DNA and associated proteins) has traditionally been studied by live cell microscopy of nuclear proteins, single genes, subchromosomal foci and chromosomal territories [Marshall et al. 1997; Belmont et al. 1998; Levi et al. 2005; Kumaran et al. 2008; Stixova et al. 2011]. Such studies are highly informative, but in practice one can only investigate a few sites simultaneously. Approaches that provide a picture of an overall chromatin dynamics have been slow to develop [Abney et al. 1997]. In the proposed research we will use our newly developed method of velocity correlation spectroscopy (VCS) [Zidovska et al. 2012a], combined with established methods like particle image velocimetry (PIV) and spatio-temporal image correlation spectroscopy (STICS), to measure high-resolution chromatin velocity maps and spatio-temporal evolution of chromatin dynamics over an entire nucleus for the first time (aim 1). In previous work using VCS we found local coherence in spatio-temporal chromatin dynamics, showing that chromatin dynamics is correlated over micron and second scale [Zidovska et al. 2012b]. Our preliminary studies also showed that chromatin dynamics uses ATP, but is independent of the cytoskeleton, hinting at nuclear ATPases being responsible for the observed dynamics. We will probe the origin of the observed chromatin dynamics by specifically inhibiting major nuclear ATPases (polymerase II, DNA polymerase and topoisomerase) using molecular poisons or RNA interference and analyzing the spatio-temporal changes in the pattern of measured chromatin dynamics (aim 2). An exciting preliminary finding showed that chromatin dynamics are largely blocked by a recently-described small molecule JQ1, which binds specifically to four BET family bromodomain proteins, antagonizing their binding to histones [Filippakopoulos et al. 2010]. By investigating how JQ1 blocks chromatin motility we will gain new information on the role of BET proteins in chromatin dynamics. Since JQ1 has demonstrated efficacy in translational models of poorly differentiated carcinoma, multiple myeloma and acute leukemia [Delmore et al. 2011; Zuber et al. 2011], this work may also help elucidate the cellular effects of a promising anti-cancer agent. Finally, we extend this approach to a small panel of cancer and non-cancer cell lines (aim 3) to ask whether chromatin dynamics are perturbed in cancer. If so, VCS analysis might prove useful as a diagnostic tool. Our discoveries in this project will provide a framework for a mechanistic picture of the origins and functional importance of chromatin dynamics in mammalian cells.

项目总结/文摘