Condensin-mediated genome organization and transcriptional regulation

凝缩蛋白介导的基因组组织和转录调控

基本信息

批准号：
9114628
负责人：
Michelle S Longworth
金额：
$ 29.83万
依托单位：
CLEVELAND CLINIC LERNER COM-CWRU
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-08-01 至 2017-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9114628
关键词：
Address Adult Affect Automobile Driving Binding Binding Sites Biological Assay Brain Cell Nucleus Cell physiology Cells ChIP-seq Characteristics Chromatin Chromatin Loop Chromatin Structure Chromosome Structures Chromosome Territory Chromosomes Co-Immunoprecipitations Color Complex Coupled DNA DNA Sequence Data Defect Development Double-Stranded RNA Drosophila genus Drosophila melanogaster Elements Event Family Gene Cluster Gene Expression Genes Genetic Materials Genetic Transcription Genome Genomic Instability Goals Health High-Throughput Nucleotide Sequencing Higher Order Chromatin Structure Human Immunofluorescence Immunologic In Vitro Interphase Knowledge Lead Life Link Malignant Neoplasms Mass Spectrum Analysis Mediating Mitosis Multiprotein Complexes Nuclear Nuclear Lamina Nuclear Pore Organism Orthologous Gene Physical condensation Positioning Attribute Proteins Regulation Research Role Syndrome Testing Therapeutic Intervention Tissues Transcript Transcriptional Regulation Transgenic Organisms Work Yeasts condensin fly human disease in vivo interest knock-down member mutant novel programs research study tumor

项目摘要

DESCRIPTION (provided by applicant): Project Summary The reorganization of specific loci within the nucleus is often tightly coupled with the timely activation of specific programs of transcription, but the complexes driving these changes are not well characterized and mechanisms that regulate these events are unknown. Efficient organization and condensation of chromatin at the beginning of mitosis requires a conserved family of complexes called Condensins. Our research has uncovered a novel role for the Drosophila Condensin II subunit, dCAP-D3 in regulating the transcription of clusters of functionally-related genes. Our long-term goal is to identify the mechanisms by which the organization and positioning of the global chromatin structure helps to coordinate local transcription events in vivo. Identification of dCAP-D3 direct targets and sequences necessary for dCAP-D3/Condensin II mediated gene cluster regulation will be accomplished through ChIP- seq analyses followed by comparison to our previous microarray data. Analysis of gene transcription in transgenic lines deficient for dCAP-D3 binding sites at target gene clusters will also be performed. To identify associated proteins necessary for dCAP-D3's ability to regulate gene clusters in vivo, dsRNAs will be used to deplete Condensin II subunits in vivo and then test whether transcript levels of dCAP-D3 regulated gene clusters are affected. IP/Mass spectrometry analyses will identify novel in vivo dCAP-D3 binding partners and their contributions to dCAP-D3-mediated transcriptional regulation of gene clusters will also be studied. The higher order chromatin structure and sub-nuclear localization of dCAP-D3 regulated gene clusters will be examined by performing assays which detect DNA looping events and two color DNA-FISH combined with immunofluorescence. The contribution of the proposed research will be to provide previously unknown, mechanistic links between the global organization of chromatin domains and local transcriptional control. This contribution will be significant because it will further our understanding of the three-dimensional aspects of transcriptional regulation that occur inside a living organism. Since our preliminary data suggests that the ability of CAP-D3 to regulate gene clusters is conserved in human cells, mechanisms of dCAP-D3-mediated transcriptional regulation might also be conserved. Misregulation of global chromatin organizers is implicated in developmental syndromes and cancers. Therefore, elucidation of the mechanisms by which dCAP-D3/Condensin II regulates transcription will potentially uncover new avenues for therapeutic intervention in a number of human diseases.

描述（由申请人提供）：项目摘要细胞核内特定基因座的重组通常与特定转录程序的及时激活紧密相关，但驱动这些变化的复合物尚未得到很好的表征，调节这些事件的机制也未知。有丝分裂开始时染色质的有效组织和浓缩需要一个称为浓缩蛋白的保守复合物家族。我们的研究发现了果蝇凝缩蛋白 II 亚基 dCAP-D3 在调节功能相关基因簇转录中的新作用。我们的长期目标是确定整体染色质结构的组织和定位有助于协调体内局部转录事件的机制。 dCAP-D3 直接靶标和 dCAP-D3/Condensin II 介导的基因簇调控所需序列的鉴定将通过 ChIP-seq 分析来完成，然后与我们之前的微阵列数据进行比较。还将对目标基因簇上缺乏 dCAP-D3 结合位点的转基因系中的基因转录进行分析。为了鉴定 dCAP-D3 体内调节基因簇能力所需的相关蛋白，将使用 dsRNA 消耗体内 Condensin II 亚基，然后测试 dCAP-D3 调节基因簇的转录水平是否受到影响。 IP/质谱分析将鉴定新型体内 dCAP-D3 结合伴侣，并且还将研究它们对 dCAP-D3 介导的基因簇转录调控的贡献。 dCAP-D3 调节基因簇的高级染色质结构和亚核定位将通过检测 DNA 环事件和两色 DNA-FISH 与免疫荧光相结合的检测来检查。拟议研究的贡献将是提供染色质结构域的全局组织和局部转录控制之间以前未知的机制联系。这一贡献将是意义重大的，因为它将进一步加深我们对生物体内发生的转录调控的三维方面的理解。由于我们的初步数据表明 CAP-D3 调节基因簇的能力在人类细胞中是保守的，因此 dCAP-D3 介导的转录调节机制也可能是保守的。整体染色质组织者的失调与发育综合征和癌症有关。因此，阐明 dCAP-D3/Condensin II 调节转录的机制将有可能为许多人类疾病的治疗干预开辟新途径。

项目成果

期刊论文数量（3）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Crossing the LINE Toward Genomic Instability: LINE-1 Retrotransposition in Cancer.

跨越基因组不稳定的界限：癌症中的 LINE-1 逆转录转座。

DOI：
发表时间：
2015
期刊：
影响因子：
0
作者：
Kemp, Jacqueline R;Longworth, Michelle S
通讯作者：
Longworth, Michelle S

Condensin II subunit dCAP-D3 restricts retrotransposon mobilization in Drosophila somatic cells.

凝缩蛋白 II 亚基 dCAP-D3 限制果蝇体细胞中逆转录转座子的动员。

DOI：
发表时间：
2013-10
期刊：
影响因子：
4.5
作者：
Schuster, Andrew T;Sarvepalli, Kavitha;Murphy, Eain A;Longworth, Michelle S
通讯作者：
Longworth, Michelle S

Drosophila Condensin II subunit Chromosome-associated protein D3 regulates cell fate determination through non-cell-autonomous signaling.

果蝇凝缩蛋白 II 亚基染色体相关蛋白 D3 通过非细胞自主信号传导调节细胞命运决定。

DOI：
发表时间：
2016-08-01
期刊：
影响因子：
0
作者：
Klebanow, Lindsey R;Peshel, Emanuela C;Schuster, Andrew T;De, Kuntal;Sarvepalli, Kavitha;Lemieux, Madeleine E;Lenoir, Jessica J;Moore, Adrian W;McDonald, Jocelyn A;Longworth, Michelle S
通讯作者：
Longworth, Michelle S