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5-甲基脱氧胞苷在细胞核和纯化DNA的葡萄球菌核酸酶消化产物中的分布。

Distribution of 5-methyldeoxycytidine in products of staphylococcal nuclease digestion of nuclei and purified DNA.

作者信息

Barr F G, Kastan M B, Lieberman M W

出版信息

Biochemistry. 1985 Mar 12;24(6):1424-8. doi: 10.1021/bi00327a021.

DOI:10.1021/bi00327a021
PMID:3986183
Abstract

We have compared the distribution of 5-methyldeoxycytidine (m5dC) between staphylococcal nuclease (SN) sensitive and resistant regions of human diploid fibroblast chromatin to the corresponding distribution in purified DNA. After SN digestion of fibroblast nuclei or purified DNA, nuclease-resistant products were separated from sensitive products by perchloric acid or ethanol precipitation; the radioactively labeled nucleosides were then fractionated by high-performance liquid chromatography and quantitated. Our results indicate that m5dC is preferentially associated with SN-resistant regions of both chromatin and purified DNA. The magnitudes of these preferences in fibroblast chromatin and DNA are similar; we find that the enrichment of m5dC content in SN-resistant fractions of nuclei and DNA relative to the corresponding sensitive fractions is approximately 2-3-fold. Therefore, highly methylated regions of DNA have an intrinsic resistance to digestion by SN that is of sufficient magnitude to explain the high degree of nuclease resistance of chromatin containing highly methylated DNA.

摘要

我们已经将人类二倍体成纤维细胞染色质中对葡萄球菌核酸酶（SN）敏感和抗性区域之间的5-甲基脱氧胞苷（m5dC）分布与纯化DNA中的相应分布进行了比较。在用SN消化成纤维细胞核或纯化DNA后，通过高氯酸或乙醇沉淀将核酸酶抗性产物与敏感产物分离；然后通过高效液相色谱对放射性标记的核苷进行分级分离并定量。我们的结果表明，m5dC优先与染色质和纯化DNA的SN抗性区域相关联。成纤维细胞染色质和DNA中这些偏好的程度相似；我们发现，相对于相应的敏感级分，细胞核和DNA的SN抗性级分中m5dC含量的富集约为2-3倍。因此，DNA的高度甲基化区域对SN消化具有内在抗性，其程度足以解释含有高度甲基化DNA的染色质对核酸酶的高度抗性。

相似文献

1
Distribution of 5-methyldeoxycytidine in products of staphylococcal nuclease digestion of nuclei and purified DNA.5-甲基脱氧胞苷在细胞核和纯化DNA的葡萄球菌核酸酶消化产物中的分布。
Biochemistry. 1985 Mar 12;24(6):1424-8. doi: 10.1021/bi00327a021.
2
Undermethylation of DNA in mononucleosomes solubilized by micrococcal nuclease digestion of HeLa cell nuclei.通过微球菌核酸酶消化HeLa细胞核而溶解的单核小体中DNA的甲基化不足。
Biochem Int. 1984 Aug;9(2):251-8.
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Cleavage of DNA in nuclei and chromatin with staphylococcal nuclease.用葡萄球菌核酸酶切割细胞核和染色质中的DNA。
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A comparison of the digestion of nuclei and chromatin by staphylococcal nuclease.葡萄球菌核酸酶对细胞核和染色质消化作用的比较。
Biochemistry. 1975 Jul;14(13):2915-20. doi: 10.1021/bi00684a019.

引用本文的文献

1
Genomic hypomethylation and far-5' sequence alterations are associated with carcinogen-induced activation of the hamster thymidine kinase gene.基因组低甲基化和远5'端序列改变与致癌物诱导的仓鼠胸苷激酶基因激活有关。
Mol Cell Biol. 1986 Sep;6(9):3023-33. doi: 10.1128/mcb.6.9.3023-3033.1986.
2
Analysis of the rearrangements associated with carcinogen-induced activation of the hamster thymidine kinase gene.与致癌物诱导的仓鼠胸苷激酶基因激活相关的重排分析。
Nucleic Acids Res. 1990 Jan 11;18(1):129-35. doi: 10.1093/nar/18.1.129.
3
Structural analysis of a carcinogen-induced genomic rearrangement event.
致癌物诱导的基因组重排事件的结构分析
Proc Natl Acad Sci U S A. 1992 Feb 1;89(3):942-6. doi: 10.1073/pnas.89.3.942.