European Molecular Biology Laboratory, Heidelberg, Germany.
Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, CA, USA.
Nat Struct Mol Biol. 2023 Aug;30(8):1216-1223. doi: 10.1038/s41594-023-01008-5. Epub 2023 Jun 8.
Subnuclear compartmentalization has been proposed to play an important role in gene regulation by segregating active and inactive parts of the genome in distinct physical and biochemical environments. During X chromosome inactivation (XCI), the noncoding Xist RNA coats the X chromosome, triggers gene silencing and forms a dense body of heterochromatin from which the transcription machinery appears to be excluded. Phase separation has been proposed to be involved in XCI, and might explain the exclusion of the transcription machinery by preventing its diffusion into the Xist-coated territory. Here, using quantitative fluorescence microscopy and single-particle tracking, we show that RNA polymerase II (RNAPII) freely accesses the Xist territory during the initiation of XCI. Instead, the apparent depletion of RNAPII is due to the loss of its chromatin stably bound fraction. These findings indicate that initial exclusion of RNAPII from the inactive X reflects the absence of actively transcribing RNAPII, rather than a consequence of putative physical compartmentalization of the inactive X heterochromatin domain.
亚核隔室化被认为在基因调控中发挥重要作用,它将基因组的活性和非活性部分分隔在不同的物理和生化环境中。在 X 染色体失活(XCI)过程中,非编码的 Xist RNA 包裹 X 染色体,触发基因沉默,并形成致密的异染色质体,转录机制似乎被排除在外。相分离被认为参与了 XCI,并可能通过阻止其扩散到 Xist 包裹的区域来解释转录机制的排除。在这里,我们使用定量荧光显微镜和单粒子跟踪技术,显示 RNA 聚合酶 II(RNAPII)在 XCI 起始时可自由进入 Xist 区域。相反,RNAPII 的表观耗竭是由于其与染色质稳定结合的部分丢失。这些发现表明,初始时 RNAPII 从失活 X 染色体中的排除反映了活跃转录的 RNAPII 的缺失,而不是失活 X 染色体异染色质域的假定物理隔室化的结果。
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