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[通过流式细胞术研究哺乳动物细胞核中的染色质包装]

[Chromatin packing in the nuclei of mammalian cells studied by flow cytometry].

作者信息

Filatov M V, Varfolomeeva E Iu

出版信息

Tsitologiia. 1990;32(4):343-51.

PMID:1700520
Abstract

A phenomenon of limited accessibility of mammalian cell nuclear DNA to external agents is described, which is revealed by altered binding of DNA-specific fluorescent dyes after fixation of nuclei with glutaraldehyde. This approach is demonstrated to permit visualizing different degrees of DNA shielding by proteins in cells of diverse differentiation and functional state, as well as measuring an alteration of nature chromatin conformation after different treatment. The treatment of nuclei with single-stranded DNA was found to result in a considerable increase in nuclear DNA accessibility to fluorescent dyes, probably by activation of endogenous nuclear DNA-dependent serine proteases. The data obtained allow to suppose the existence of a specific nuclear DNA accessibility regulating system based on protease action and regulated by the appearance of single-stranded DNA stretches which accompanied such genetic processes as DNA repair, transcription, recombination and replication.

摘要

描述了一种哺乳动物细胞核DNA对外部试剂可及性有限的现象,在用戊二醛固定细胞核后,DNA特异性荧光染料结合的改变揭示了这一现象。已证明这种方法能够可视化不同分化和功能状态细胞中蛋白质对DNA的不同程度屏蔽,以及测量不同处理后天然染色质构象的改变。发现用单链DNA处理细胞核会导致细胞核DNA对荧光染料的可及性显著增加,这可能是通过激活内源性核DNA依赖性丝氨酸蛋白酶实现的。所获得的数据使人们能够推测存在一种基于蛋白酶作用且受单链DNA片段出现调节的特异性核DNA可及性调节系统,单链DNA片段伴随着DNA修复、转录、重组和复制等遗传过程出现。

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