Oberbeckmann Elisa, Oudelaar A Marieke
Department of Molecular Biology, Max Planck Institute for Multidisciplinary Sciences, Am Fassberg 11, 37077 Göttingen, Germany.
Genome Organization and Regulation, Max Planck Institute for Multidisciplinary Sciences, Am Fassberg 11, 37077 Göttingen, Germany.
Biochem Soc Trans. 2024 Apr 24;52(2):793-802. doi: 10.1042/BST20230883.
Eukaryotic genomes are compacted and organized into distinct three-dimensional (3D) structures, which range from small-scale nucleosome arrays to large-scale chromatin domains. These chromatin structures play an important role in the regulation of transcription and other nuclear processes. The molecular mechanisms that drive the formation of chromatin structures across scales and the relationship between chromatin structure and function remain incompletely understood. Because the processes involved are complex and interconnected, it is often challenging to dissect the underlying principles in the nuclear environment. Therefore, in vitro reconstitution systems provide a valuable approach to gain insight into the molecular mechanisms by which chromatin structures are formed and to determine the cause-consequence relationships between the processes involved. In this review, we give an overview of in vitro approaches that have been used to study chromatin structures across scales and how they have increased our understanding of the formation and function of these structures. We start by discussing in vitro studies that have given insight into the mechanisms of nucleosome positioning. Next, we discuss recent efforts to reconstitute larger-scale chromatin domains and loops and the resulting insights into the principles of genome organization. We conclude with an outlook on potential future applications of chromatin reconstitution systems and how they may contribute to answering open questions concerning chromatin architecture.
真核生物基因组被压缩并组织成不同的三维（3D）结构，其范围从小规模的核小体阵列到大规模的染色质结构域。这些染色质结构在转录调控和其他核过程中发挥着重要作用。驱动跨尺度染色质结构形成的分子机制以及染色质结构与功能之间的关系仍未完全了解。由于所涉及的过程复杂且相互关联，在核环境中剖析其潜在原理往往具有挑战性。因此，体外重建系统为深入了解染色质结构形成的分子机制以及确定所涉及过程之间的因果关系提供了一种有价值的方法。在本综述中，我们概述了用于研究跨尺度染色质结构的体外方法，以及这些方法如何增进我们对这些结构形成和功能的理解。我们首先讨论了深入了解核小体定位机制的体外研究。接下来，我们讨论了重建更大规模染色质结构域和环的最新努力以及由此对基因组组织原理的见解。我们最后展望了染色质重建系统未来可能的应用，以及它们如何有助于回答有关染色质结构的开放性问题。
