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在非洲爪蟾卵母细胞核提取物中组装间距正确的染色质。

Assembly of correctly spaced chromatin in a nuclear extract from Xenopus laevis oocytes.

作者信息

Sessa G, Ruberti I

机构信息

Dipartimento di Genetica e Biologia Molecolare, Università di Roma La Sapienza, Italy.

出版信息

Nucleic Acids Res. 1990 Sep 25;18(18):5449-55. doi: 10.1093/nar/18.18.5449.

DOI:10.1093/nar/18.18.5449
PMID:2170936
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC332223/
Abstract

Assembly of nucleosomes on relaxed, covalently closed DNA has been studied in a nuclear extract of Xenopus laevis oocytes. Nucleosomes containing the four histones H3, H4, H2A and H2B but lacking histone H1 are readily assembled on the DNA. The pattern of micrococcal nuclease digestion shows that the nucleosomes assembled in the absence of ATP and Mg (II) are closely packed, with a periodicity of 150 base pairs (bp). In contrast, in the presence of ATP and Mg (II) the spacing of the nucleosomes is 180 bp, similar to that observed for nucleosomes assembled on DNA microinjected into oocyte nuclei. The ATP and Mg (II) requirements for the assembly of correctly spaced nucleosomes are unrelated to the activity of the ATP and Mg (II) dependent DNA topoisomerase II in the extract; addition of specific inhibitors of eukaryotic DNA topoisomerase II has no effect on the spacing of the reconstituted nucleosomes. The ATP requirement in the assembly of correctly spaced nucleosomes can be substituted by adenosine 5'-O-3'-thiotriphosphate (gamma-S-ATP) but not by adenyl-5'-yl imidodiphosphate (AMP-P-(NH)-P).

摘要

已在非洲爪蟾卵母细胞的核提取物中研究了松弛的、共价闭合的DNA上核小体的组装。含有组蛋白H3、H4、H2A和H2B但缺乏组蛋白H1的核小体很容易在DNA上组装。微球菌核酸酶消化模式表明，在没有ATP和Mg（II）的情况下组装的核小体紧密堆积，周期为150个碱基对（bp）。相比之下，在有ATP和Mg（II）的情况下，核小体的间距为180 bp，类似于在注入卵母细胞核的DNA上组装的核小体所观察到的间距。正确间隔的核小体组装对ATP和Mg（II）的需求与提取物中ATP和Mg（II）依赖性DNA拓扑异构酶II的活性无关；添加真核DNA拓扑异构酶II的特异性抑制剂对重组核小体的间距没有影响。正确间隔的核小体组装中对ATP的需求可以被腺苷5'-O-3'-硫代三磷酸（γ-S-ATP）替代，但不能被腺苷-5'-基亚氨基二磷酸（AMP-P-(NH)-P）替代。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/26f6/332223/a7581cd579e1/nar00202-0110-c.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/26f6/332223/e55d3b82dbd1/nar00202-0108-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/26f6/332223/d27f09b0326f/nar00202-0108-b.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/26f6/332223/89305f4bd5a6/nar00202-0109-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/26f6/332223/d965d7249c7f/nar00202-0109-c.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/26f6/332223/9e4a0f1eda7e/nar00202-0110-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/26f6/332223/82d1ad5cea0b/nar00202-0110-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/26f6/332223/a7581cd579e1/nar00202-0110-c.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/26f6/332223/e55d3b82dbd1/nar00202-0108-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/26f6/332223/d27f09b0326f/nar00202-0108-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/26f6/332223/fbddc73d5529/nar00202-0109-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/26f6/332223/89305f4bd5a6/nar00202-0109-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/26f6/332223/d965d7249c7f/nar00202-0109-c.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/26f6/332223/9e4a0f1eda7e/nar00202-0110-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/26f6/332223/82d1ad5cea0b/nar00202-0110-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/26f6/332223/a7581cd579e1/nar00202-0110-c.jpg

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引用本文的文献

1
DNA-dependent phosphorylation of histone H2A.X during nucleosome assembly in Xenopus laevis oocytes: involvement of protein phosphorylation in nucleosome spacing.非洲爪蟾卵母细胞中核小体组装过程中组蛋白H2A.X的DNA依赖性磷酸化：蛋白质磷酸化在核小体间距中的作用。
EMBO J. 1991 Oct;10(10):3043-50. doi: 10.1002/j.1460-2075.1991.tb07855.x.

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