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以不同速率转录的基因在染色质中具有相似的构象。

Genes transcribed at diverse rates have a similar conformation in chromatin.

作者信息

Garel A, Zolan M, Axel R

出版信息

Proc Natl Acad Sci U S A. 1977 Nov;74(11):4867-71. doi: 10.1073/pnas.74.11.4867.

Abstract

We have analyzed the DNA generated upon treatment of oviduct nuclei with pancreatic DNase I (deoxyribonucleate 3'-oligonucleotidohydrolase; EC 3.1.4.6), with cDNA copies of specific mRNA sequences to study the structure and organization of transcriptionally active genes in chromatin. In this report we examine the kinetics of digestion of three classes of genes in the oviduct which are transcribed at significantly different rates. Our results indicate that the ovalbumin genes appear to be organized by chromatin proteins in such a way that they are rendered exceedingly sensitive to digestion by DNase I. This sensitivity is not observed in the liver, a tissue in which these genes are transcriptionally inert. Furthermore, the transcriptionally inactive globin genes in the oviduct are not selectively sensitive to nuclease attack and are digested 5 times more slowly in the ovalbumin genes in this tissue. In addition, we have examined the accessibility of a complex subset of genes that are rarely represented in the mRNA and are likely to be transcribed at a frequency orders of magnitude below that of the ovalbumin gene. Comparison of the accessibility of these sequences with that of the ovalbumin gene indicates that these two subsets of genes are recognized and cleaved by DNase I at similar rates. These results suggest that the maintenance of an active conformation about specific genes does not reflect the polymerase distribution about these genes. This active conformation is therefore not confined to sequences actively engaged in the transcription process and may reflect the structure about a subpopulation of the genome which represents the transcriptional potential of a given cell type.

摘要

我们分析了用胰脱氧核糖核酸酶I(脱氧核糖核酸3'-寡核苷酸水解酶;EC 3.1.4.6)处理输卵管细胞核后产生的DNA,并利用特定mRNA序列的cDNA拷贝来研究染色质中转录活性基因的结构和组织。在本报告中,我们研究了输卵管中三类转录速率差异显著的基因的消化动力学。我们的结果表明,卵清蛋白基因似乎是以一种使其对脱氧核糖核酸酶I的消化异常敏感的方式由染色质蛋白组织起来的。在肝脏中未观察到这种敏感性,在肝脏中这些基因是转录惰性的。此外,输卵管中转录无活性的珠蛋白基因对核酸酶攻击没有选择性敏感性,在该组织中其被消化的速度比卵清蛋白基因慢5倍。另外,我们研究了一组复杂的基因的可及性,这些基因在mRNA中很少出现,其转录频率可能比卵清蛋白基因低几个数量级。将这些序列的可及性与卵清蛋白基因的可及性进行比较表明,这两组基因被脱氧核糖核酸酶I识别和切割的速率相似。这些结果表明,特定基因活性构象的维持并不反映这些基因周围的聚合酶分布。因此,这种活性构象并不局限于积极参与转录过程的序列,可能反映了基因组中代表给定细胞类型转录潜能的一个亚群的结构。

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Nuclease sensitivity of active chromatin.活性染色质的核酸酶敏感性
Nucleic Acids Res. 1980 Nov 25;8(22):5143-55. doi: 10.1093/nar/8.22.5143.

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Chromatin accessibility: a window into the genome.染色质可及性:窥探基因组的窗口。
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Proc Natl Acad Sci U S A. 1981 Jul;78(7):4246-50. doi: 10.1073/pnas.78.7.4246.

本文引用的文献

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Animal DNA-dependent RNA polymerases. 12. Determination of the cellular number of RNA polymerase B molecules.
Biochim Biophys Acta. 1974 Jun 27;353(2):185-92. doi: 10.1016/0005-2787(74)90183-x.
4
The in vitro synthesis of avian myeloblastosis viral RNA sequences.禽成髓细胞瘤病毒RNA序列的体外合成
Proc Natl Acad Sci U S A. 1974 Aug;71(8):3045-9. doi: 10.1073/pnas.71.8.3045.
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Transcription in polytene chromosomes.
Cell. 1975 Jan;4(1):1-9. doi: 10.1016/0092-8674(75)90127-0.
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Structure of transcriptionally active chromatin.转录活性染色质的结构。
Proc Natl Acad Sci U S A. 1975 Nov;72(11):4404-8. doi: 10.1073/pnas.72.11.4404.

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