Zhi Yang Tan, Shujun Cai, Saayli A Paithankar, Tingsheng Liu, Xin Nie, Jian Shi, Lu Gan
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引用次数: 0
摘要
当缺乏氮元素时,裂殖酵母细胞会进入静止的 "G0 "状态,细胞核变小,转录抑制。庞贝酵母 G0 细胞的基因组学已经得到了很好的研究,但其核细胞生物学的大部分内容仍然未知。在这里,我们使用共聚焦显微镜、免疫印迹和电子冷冻成像技术研究了 S. pombe 增殖细胞、G1-停滞细胞和 G0 细胞核在细胞学、生物化学和超微结构方面的差异,重点研究了组蛋白乙酰化、RNA 聚合酶 II 的命运和大分子复合物的包装。与增殖细胞相比,G0 细胞的组蛋白乙酰化、核 RNA 聚合酶 II 和活跃转录水平较低。G0细胞核具有类似的大分子拥挤现象,但染色质相关的多质子球状复合物较少。在氮饥饿过程中诱导组蛋白过乙酰化会导致细胞核变大,从而使染色质不那么紧密。然而,这些组蛋白过乙酰化的细胞仍然受到转录抑制,具有类似的核拥挤现象。在增殖细胞、G1-停滞细胞和 G0 细胞中,典型核小体(与晶体结构相似的核小体)很少见。因此,我们的研究表明,细胞核生理发生极端变化时,大分子水平不会发生极端重组。
Macromolecular and cytological changes in fission yeast G0 nuclei.
When starved of nitrogen, fission yeast Schizosaccharomyces pombe cells enter a quiescent "G0" state with smaller nuclei and transcriptional repression. The genomics of S. pombe G0 cells has been well studied, but much of its nuclear cell biology remains unknown. Here we use confocal microscopy, immunoblots, and electron cryotomography to investigate the cytological, biochemical, and ultrastructural differences between S. pombe proliferating, G1-arrested, and G0 cell nuclei, with an emphasis on the histone acetylation, RNA polymerase II fates, and macromolecular complex packing. Compared to proliferating cells, G0 cells have lower levels of histone acetylation, nuclear RNA polymerase II, and active transcription. The G0 nucleus has similar macromolecular crowding yet fewer chromatin-associated multi-megadalton globular complexes. Induced histone hyperacetylation during nitrogen starvation results in cells that have larger nuclei and therefore less compact chromatin. However, these histone-hyperacetylated cells remain transcriptionally repressed with similar nuclear crowding. Canonical nucleosomes - those that resemble the crystal structure - are rare in proliferating, G1-arrested, and G0 cells. Our study therefore shows that extreme changes in nucleus physiology are possible without extreme reorganisation at the macromolecular level.