Characterizing the RNA-mediated recruitment of histone deacetylases to chromatin

表征 RNA 介导的组蛋白脱乙酰酶向染色质的募集

基本信息

批准号：
10680276
负责人：
James Kang Guo
金额：
$ 5.27万
依托单位：
CALIFORNIA INSTITUTE OF TECHNOLOGY
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-06-01 至 2028-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10680276
关键词：
Affinity Chromatography Antisense RNA Binding Binding Proteins Binding Sites Cancerous Cell Cycle Cell Nucleus Cells Chromatin Complex DNA Binding DNA-Binding Proteins Data Enzymes Future Gene Expression Gene Expression Regulation Gene Silencing Gene Targeting Genes Genetic Genome Guide RNA HDAC1 gene HDAC3 gene Health Histone Deacetylase Histone Deacetylase Inhibitor Histone Deacetylation Human Immunoprecipitation Knock-out Link Malignant Neoplasms Mass Spectrum Analysis Measures Mediating Methods Mission Molecular Multiprotein Complexes Mutate Nuclear Pathology Patients Pharmaceutical Preparations Protein Isoforms Proteins RNA RNA Binding RNA-Binding Proteins Repression Research Research Project Grants Role Site Specificity Suppressor-Effector T-Lymphocytes Testing Therapeutic Time Toxic effect Tumor Suppressor Genes United States National Institutes of Health Untranslated RNA Work X Chromosome cancer therapy cancer type chromatin modification crosslink effective therapy experimental study gene repression genetic manipulation genetic regulatory protein genomic locus improved in vivo inhibitor link protein mRNA Precursor novel pharmacologic protein complex public database recruit side effect therapeutic target transcription factor tumor

Affinity Chromatography Antisense RNA Binding Binding Proteins Binding Sites Cancerous Cell Cycle Cell Nucleus Cells Chromatin Complex DNA Binding DNA-Binding Proteins Data Enzymes Future Gene Expression Gene Expression Regulation Gene Silencing Gene Targeting Genes Genetic Genome Guide RNA HDAC1 gene HDAC3 gene Health Histone Deacetylase Histone Deacetylase Inhibitor Histone Deacetylation Human Immunoprecipitation Knock-out Link Malignant Neoplasms Mass Spectrum Analysis Measures Mediating Methods Mission Molecular Multiprotein Complexes Mutate Nuclear Pathology Patients Pharmaceutical Preparations Protein Isoforms Proteins RNA RNA Binding RNA-Binding Proteins Repression Research Research Project Grants Role Site Specificity Suppressor-Effector T-Lymphocytes Testing Therapeutic Time Toxic effect Tumor Suppressor Genes United States National Institutes of Health Untranslated RNA Work X Chromosome cancer therapy cancer type chromatin modification crosslink effective therapy experimental study gene repression genetic manipulation genetic regulatory protein genomic locus improved in vivo inhibitor link protein mRNA Precursor novel pharmacologic protein complex public database recruit side effect therapeutic target transcription factor tumor

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项目摘要

PROJECT SUMMARY Global histone deacetylation is linked to many types of cancer and is controlled by histone deacetylases (HDACs). Although HDAC inhibitors are widely used in cancer treatment, their activity does not target specific HDAC isoforms nor specific genes and therefore results in significant side effects for patients. Thus, there is a pressing need to target HDAC activity in a highly precise, gene-specific manner to develop safer and more effective treatments. The overall objective of this proposal is to understand how HDACs are recruited to specific genome regions. Because HDACs do not possess intrinsic DNA binding activity, they are thought to be recruited to chromatin through interactions with DNA binding proteins, though this mechanism has not been fully explored. Recently, our lab and others identified that the SHARP RNA-binding protein directly interacts with the Xist long noncoding RNA (lncRNA) to specifically recruit HDAC3 to the future inactive X chromosome (Xi). In this way, the Xist-SHARP/SMRT/HDAC3 repressive complex deacetylates histones and silences gene expression on the Xi. Our lab has also demonstrated that SHARP localizes to many nuclear sites (beyond the Xi) in an RNA-dependent manner, raising the question of which additional RNAs recruit it (along with HDAC3) and to what specific genomic locations. Notably, nearly all human HDACs associate within multi-protein complexes containing SHARP and other RNA-binding proteins, suggesting that this mechanism of RNA-guidance may extend beyond SHARP and Xist. I hypothesize that HDAC complexes are recruited by RNAs to achieve specificity to their various regulatory targets throughout the nucleus. First, I will determine which SHARP-RNA interactions are functionally important by genetically perturbing the SHARP binding sites of candidate RNAs. I will then measure effects on gene expression and if HDAC3 activity is required for these effects (Aim 1). Second, I will comprehensively identify RNA-binding proteins associated with other cancer-associated HDACs and define their in vivo RNA binding sites. I will then determine which protein-bound candidate RNAs from this screen are regulated in an HDAC-dependent manner (Aim 2). The proposed research has the potential to transform our understanding of RNAs and RNA- binding proteins as central regulators in organizing chromatin modifications by HDACs in both normal and cancerous cell states.

项目摘要全球组蛋白脱乙酰化与多种类型的癌症有关，由组蛋白脱乙酰基酶控制（HDACS）。尽管HDAC抑制剂被广泛用于癌症治疗，但它们的活性并不针对特定 HDAC同工型或特定基因，因此会对患者产生重大副作用。因此，有一个紧迫需要以高度精确的基因特异性方式靶向HDAC活动，以发展更安全和更多有效的治疗方法。该提案的总体目的是了解如何将HDAC招募到特定基因组区域。由于HDAC不具有固有的DNA结合活性，因此被认为是招募的通过与DNA结合蛋白相互作用的染色质，尽管尚未完全探索该机制。最近，我们的实验室和其他实验室发现尖锐的RNA结合蛋白直接与Xist长相互作用非编码RNA（LNCRNA）可将HDAC3专门募集到未来的无活性X染色体（XI）中。这样， XIST-SHARP/SMRT/HDAC3抑制性复合物脱乙酰基，Xi上的组蛋白和沉默基因表达。我们的实验室还证明，尖锐的位置位于RNA依赖性的许多核位点（超越XI）方式，提出了一个额外RNA募集它的问题（与HDAC3一起招募）以及哪些特定基因组位置。值得注意的是，几乎所有的人类HDAC都在包含尖锐和其他RNA结合蛋白，表明这种RNA戒断机制可能延伸到尖锐和 xist。我假设RNA招募了HDAC复合物，以实现其各种监管的特异性整个核的目标。首先，我将确定哪些尖锐的RNA相互作用在功能上很重要通过遗传扰动候选RNA的尖锐结合位点。然后，我将测量对基因的影响这些效果需要表达，如果需要HDAC3活性（AIM 1）。其次，我将全面识别与其他与癌症相关的HDAC相关的RNA结合蛋白，并定义其体内RNA结合位点。然后，我将在此屏幕中确定哪些蛋白质结合的候选RNA在HDAC依赖性方面受到调节方式（目标2）。拟议的研究有可能改变我们对RNA和RNA-的理解在正常和癌细胞状态。