Gottesfeld J M, Bagi G, Berg B, Bonner J
Biochemistry. 1976 Jun 1;15(11):2472-83. doi: 10.1021/bi00656a034.
Rat liver chromatin has been separated into nuclease-sensitive and -resistant fractions after mild digestion with DNAase II. The nuclease-sensitive material is further fractionated into Mg2+ -soluble and -insoluble chromatin fractions. The kinetics of production of these chromatin fractions have been investigated. After a brief enzyme treatment (5 min at 10 enzyme units/A260 unit of chromatin at pH 6.6), 11% of the input chromatin DNA is found in the Mg2+ -soluble fraction. This DNA has a weight-average single-strand length of about 400 nucleotides and, as determined by renaturation kinetics, comprises a subset of nonrepetitive DNA sequences and a subset of families of middle repetitive sequences. This demonstrates the nonrandom distribution of repetitive and single copy sequences in the Mg2+ -soluble fraction of chromatin. Previous studies have shown that the Mg2+ -soluble fraction is enriched in nonrepeated sequences which are transcribed in vivo (Gottesfeld, J.M., Garrard, W.T., Bagi, G., Wilson, R.F., and Bonner, J. (1974), Proc. Natl. Acad. Sci. U.S.A. 71, 2193-2197). We now report that the Mg2+ -soluble fraction of liver chromatin contains a low proportion of sequences in common with the Mg2+ -soluble fraction of brain chromatin. Thus, fractionation does not depend on some general property of chromatin but is specific with regard to the template activity of the tissue from which the chromatin was obtained.
用DNA酶II轻度消化后,大鼠肝脏染色质已被分离为核酸酶敏感和抗性部分。核酸酶敏感物质进一步分离为Mg2 +可溶性和不溶性染色质部分。已经研究了这些染色质部分的产生动力学。经过短暂的酶处理(在pH 6.6下以10个酶单位/A260单位染色质处理5分钟),在Mg2 +可溶性部分中发现了11%的输入染色质DNA。该DNA的重均单链长度约为400个核苷酸,通过复性动力学测定,它包含非重复DNA序列的一个子集和中度重复序列家族的一个子集。这证明了染色质Mg2 +可溶性部分中重复序列和单拷贝序列的非随机分布。先前的研究表明,Mg2 +可溶性部分富含体内转录的非重复序列(Gottesfeld,J.M.,Garrard,W.T.,Bagi,G.,Wilson,R.F.和Bonner,J.(1974年),美国国家科学院院刊71,2193 - 2197)。我们现在报告,肝脏染色质的Mg2 +可溶性部分与脑染色质的Mg2 +可溶性部分共有的序列比例较低。因此,分级分离并不取决于染色质的某些一般特性,而是对于从中获得染色质的组织的模板活性具有特异性。