Investigating the regulation of chromatin associated proteins by intrinsically disordered regions

研究本质上无序区域对染色质相关蛋白的调节

• 批准号: 2203311
• 负责人: Catherine Musselman
• 金额: $ 130万
• 依托单位: University of Colorado at Denver
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-01 至 2027-08-31
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2203311&HistoricalAwards=false
• 关键词: Investigating; regulation; chromatin; associated; proteins

Each cell in a multi-cell organism originally contains a full copy of the genome in its nucleus, but only a subset of these genes is active and in use. Proper regulation of which genes are on, and which are off, in each cell is critical to steer development and all basic body functions. The genome does not exist as naked DNA but is wrapped around complexes of proteins called histones, which together with the DNA form so-called chromatin. Gene regulation is in large part dictated by how this chromatin is packaged around individual genes, which is in turn controlled by various large protein complexes in the nucleus. A critical aspect of these protein complexes’ function is the ability to read post-translational modifications (PTMs) on the histones, which act as labels to guide them to the correct place. However, how these PTMs contribute to this process is still poorly understood within the overall complexity of chromatin. This project addresses fundamental questions of how histone PTM readout in a chromatin context is regulated by highly conserved elements within these chromatin-binding proteins to control proper gene transcription during development. In addition to the scientific goals, the PIs will work closely with a local commuter college to recruit and retain young students from underrepresented groups into the sciences. They will also work to enhance communication between the scientific and lay communities by encouraging the use of art in this conversation, through participation in the Art of Science Fellowship. The questions addressed in this project are fundamentally important for understanding how histone PTM readout is regulated in the chromatin context. Specifically, this project will investigate how evolutionarily conserved, intrinsically disordered regions (IDRs) found adjacent to histone reader domains regulate nucleosome dynamics, histone PTM readout, transcription, and development. The mechanistic basis of this regulation will be uncovered using a series of NMR spectroscopy experiments on conserved IDRs-reader pairs. This project will test the functional importance of these IDRs in vivo using an allelic series of zebrafish mutants. Selective removal and substitution of these deeply conserved amino acid residues and comparison to loss-of-function alleles will allow, for the first time, testing of the developmental and physiological contribution of these IDRs. This project was supported jointly by Molecular Biophysics and Genetic Mechanism Clusters of the Division of Molecular and Cellular Biosciences.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

多细胞生物中的每个细胞最初都包含其核中基因组的完整副本，但只有这些基因的一个子集是活性和使用的。在每个细胞中，适当调节哪些基因正在启用，哪些基因对转向发育和所有基本身体功能至关重要。该基因组并非作为裸DNA而存在，而是包裹在称为组蛋白的蛋白质的复合物周围，它们与DNA形成所谓的染色质一起。基因调节在很大程度上取决于该染色质如何包装在单个基因周围，而该基因又受细胞核中各种大型蛋白质复合物的控制。这些蛋白质复合物功能的一个关键方面是能够在组蛋白上读取翻译后修饰（PTM）的能力，这些能力是引导它们到正确位置的标签。但是，在染色质的整体复杂性中，这些PTM如何促进这一过程的贡献仍然很差。该项目解决了在这些染色质结合蛋白中高度组成的元素在染色质环境中如何调节组蛋白PTM如何调节的基本问题，以控制发育过程中适当的基因转录。除了科学目标外，PIS还将与当地的通勤学院紧密合作，以招募和留住来自代表性不足的群体的年轻学生进入科学。他们还将通过参与科学奖学金的艺术来鼓励在这次谈话中使用艺术，从而通过鼓励艺术在这次谈话中来增强科学和外行社区之间的沟通。该项目中解决的问题对于了解如何在染色质环境中调节组蛋白PTM读数至关重要。具体而言，该项目将研究与组蛋白读取器域相邻的进化构成，本质上无序的区域（IDR）如何调节核能动力学，组蛋白PTM读数，转录和发展。该调节的机械基础将使用构成的IDRS阅读器对的一系列NMR光谱实验来发现。该项目将使用一系列等位基因斑马鱼突变体来测试这些IDR在体内的功能重要性。选择性去除和取代这些深层保守的氨基酸保留率以及与功能丧失等位基因的比较，将首次允许测试这些IDR的发育和身体贡献。该项目得到了分子和细胞生物科学划分的分子生物物理学和遗传机制簇的共同支持。该奖项反映了NSF的法定任务，并通过使用基金会的智力优点和更广泛的影响来评估NSF的法定任务。

项目成果

Next-generation plasmids for transgenesis in zebrafish and beyond.

• DOI: 10.1242/dev.201531
• 发表时间: 2023-04-15
• 期刊: DEVELOPMENT
• 影响因子: 4.6
• 作者: Kemmler, Cassie L.;Moran, Hannah R.;Murray, Brooke F.;Scoresby, Aaron;Klem, John R.;Eckert, Rachel L.;Lepovsky, Elizabeth;Bertho, Sylvain;Nieuwenhuize, Susan;Burger, Sibylle;D'Agati, Gianluca;Betz, Charles;Puller, Ann-Christin;Felker, Anastasia;Ditrychova, Karolina;Botschi, Seraina;Affolter, Markus;Rohner, Nicolas;Ben Lovely, C.;Kwan, Kristen M.;Burger, Alexa;Mosimann, Christian
• 通讯作者: Mosimann, Christian

PBRM1 bromodomains associate with RNA to facilitate chromatin association.

• DOI: 10.1093/nar/gkad072
• 发表时间: 2023-05-08
• 期刊: Nucleic acids research
• 影响因子: 14.9