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转录活性基因和抑制基因的染色质结构。

Chromatin structure of transcriptionally competent and repressed genes.

作者信息

Kamakaka R T, Thomas J O

机构信息

Department of Biochemistry, University of Cambridge, UK.

出版信息

EMBO J. 1990 Dec;9(12):3997-4006. doi: 10.1002/j.1460-2075.1990.tb07621.x.

Abstract

We have compared transcriptionally competent and repressed genes with respect to their linker histone content and their ability to fold into higher-order structures. Histones were cross-linked covalently to DNA in chicken erythrocyte and oviduct nuclei by UV irradiation, and the DNA that was immunoprecipitated with anti-H1 and (for erythrocytes) anti-H5 antibodies was analysed for particular DNA sequences. None of the sequences investigated was free of H1 (H5). However, in mature erythrocytes the tissue-specific adult beta-globin gene (beta A) appears to be partially depleted of H5, and both the beta-globin gene and the H5 gene (also tissue-specific), as well as the 'housekeeping' beta-actin gene, appear to be partially depleted of H1 relative to inactive genes; in oviduct slight H1-depletion is detected on the ovalbumin gene relative to genes that are inactive in this tissue and the actin gene. Transcriptionally competent erythrocyte chromatin fragments, in contrast to inactive fragments, are unable to self-associate into 'pseudo-higher-order structures'. This is likely to be a consequence of the partial depletion of H5 and/or H1 in active chromatin, resulting in the breakdown of (probably cooperative) interactions between H5 and/or H1 molecules that otherwise mediate the assembly of pseudo-higher-order structures in vitro and a stable 30 nm chromatin filament in vivo.

摘要

我们比较了具有转录活性和受抑制的基因在连接组蛋白含量以及折叠成高阶结构能力方面的差异。通过紫外线照射,将组蛋白与鸡红细胞和输卵管细胞核中的DNA共价交联,然后用抗H1抗体(对于红细胞则用抗H5抗体)进行免疫沉淀,分析沉淀DNA中的特定DNA序列。所研究的序列均未不含H1(H5)。然而,在成熟红细胞中,组织特异性的成人β-珠蛋白基因(βA)似乎部分缺乏H5,并且β-珠蛋白基因和H5基因(也是组织特异性的)以及“管家”β-肌动蛋白基因,相对于无活性基因而言,似乎都部分缺乏H1;在输卵管中,相对于该组织中无活性的基因和肌动蛋白基因,卵清蛋白基因上检测到轻微的H1缺失。与无活性片段相比,具有转录活性红细胞染色质片段无法自组装成“假高阶结构”。这可能是由于活性染色质中H5和/或H1部分缺失的结果,导致H5和/或H1分子之间(可能是协同的)相互作用的破坏,否则这些相互作用在体外介导假高阶结构的组装以及在体内介导稳定的30nm染色质细丝的形成。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/393f/552172/10e9134db0bb/emboj00239-0194-a.jpg

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