Investigating the regulation of chromatin associated proteins by intrinsically disordered regions

多细胞生物体中的每个细胞最初在其细胞核中包含基因组的完整拷贝，但只有这些基因的一个子集是活跃的并在使用中。正确调节每个细胞中哪些基因开启，哪些基因关闭，对于指导发育和所有基本身体功能至关重要。基因组并不以裸DNA的形式存在，而是包裹在称为组蛋白的蛋白质复合物周围，组蛋白与DNA一起形成所谓的染色质。基因调控在很大程度上取决于染色质如何包装在单个基因周围，而这些基因又由细胞核中的各种大蛋白质复合物控制。这些蛋白质复合物功能的一个关键方面是能够读取组蛋白上的翻译后修饰（PTM），组蛋白作为标签将它们引导到正确的位置。然而，在染色质的整体复杂性中，这些PTM如何促进这一过程仍然知之甚少。该项目解决了在染色质背景下组蛋白PTM读出如何由这些染色质结合蛋白内的高度保守元件调节以控制发育过程中适当的基因转录的基本问题。除了科学目标，PI将与当地的通勤大学密切合作，从代表性不足的群体中招募和留住年轻学生进入科学领域。他们还将努力加强科学和外行社区之间的沟通，鼓励在这种对话中使用艺术，通过参与科学奖学金的艺术。在这个项目中解决的问题是从根本上了解组蛋白PTM读出是如何在染色质的情况下进行调节的重要。具体来说，该项目将研究如何进化保守，内在无序区域（IDR）发现邻近组蛋白阅读器域调节核小体动力学，组蛋白PTM读出，转录和发展。这种调节的机制基础将被发现使用一系列的NMR光谱实验保守的IDRs读者对。该项目将测试这些IDR在体内使用斑马鱼突变体的等位基因系列的功能的重要性。选择性去除和取代这些高度保守的氨基酸残基，并与功能丧失的等位基因进行比较，将允许首次测试这些IDR的发育和生理贡献。该项目由分子生物物理学和分子与细胞生物科学部的遗传机制集群共同支持。该奖项反映了NSF的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

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.