Structural and biochemical analyses of the nucleosome containing Komagataella pastoris histones
Structural and biochemical analyses of the nucleosome containing Komagataella pastoris histones
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含牧马人核糖体组蛋白的核小体结构和生化分析
DOI：
10.1093/jb/mvac043
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发表时间：
2022-04-29
影响因子：
2.7
通讯作者：
Kurumizaka, Hitoshi
中科院分区：
文献类型：
--
作者：
Fukushima, Yutaro;Hatazawa, Suguru;Kurumizaka, Hitoshi
Komagataella pastoris是一种甲基营养型酵母，通常用作蛋白质生产的宿主细胞。在本研究中，我们用K. pastoris组蛋白，并通过低温电镜确定核小体核心颗粒的结构。在K。在巴斯德氏酵母核小体中，组蛋白形成八聚体，DNA以左旋方式包裹在八聚体周围。与人核小体相比，pastoris核小体更容易接近。体外转录实验表明K. pastoris核小体由K. pastoris RNA聚合酶II（RNAPII）的表达效率高于人核小体，而RNAPII在K. pastoris核小体与人核小体的核小体相同。这些结果表明，DNA末端的灵活性可能会提高转录效率的核小体，但影响最小的RNAPII的核小体暂停位置。
Komagataella pastoris is a methylotrophic yeast that is commonly used as a host cell for protein production. In the present study, we reconstituted the nucleosome with K. pastoris histones and determined the structure of the nucleosome core particle by cryogenic electron microscopy. In the K. pastoris nucleosome, the histones form an octamer and the DNA is left-handedly wrapped around it. Micrococcal nuclease assays revealed that the DNA ends of the K. pastoris nucleosome are somewhat more accessible, as compared with those of the human nucleosome. In vitro transcription assays demonstrated that the K. pastoris nucleosome is transcribed by the K. pastoris RNA polymerase II (RNAPII) more efficiently than the human nucleosome, while the RNAPII pausing positions of the K. pastoris nucleosome are the same as those of the human nucleosome. These results suggested that the DNA end flexibility may enhance the transcription efficiency in the nucleosome but minimally affect the nucleosomal pausing positions of RNAPII.