Nicolas R H, Wright C A, Cockerill P N, Wyke J A, Goodwin G H
Nucleic Acids Res. 1983 Feb 11;11(3):753-72. doi: 10.1093/nar/11.3.753.
Brief micrococcal nuclease digestion of chick embryonic red blood cells results in preferential excision and solubilization of monomer nucleosomes associated with beta-globin sequences and also 5'-sequences flanking the beta-globin gene. Both regions are DNAse-I sensitive in nuclei. Such salt-soluble nucleosomes are enriched in all four major HMG proteins but HMG1 and 2 are only weakly associated. These nucleosomes appear to have lost much of the DNAse-I sensitivity of active genes. The HMG14 and 17-containing salt-soluble nucleosomes separated by electrophoresis are not DNAse-I sensitive and contain inactive gene sequences as well as active sequences. Reconstitution of HMG proteins onto bulk nucleosomes or chromatin failed to reveal an HMG-dependent sensitivity of active genes as assayed by dot-blot hybridization and it was found that the DNAse-I sensitivity of ASV proviral sequences as assayed by dot-blot hybridization was not HMG-dependent. These results indicate that higher order chromatin structures might be responsible for nuclease sensitivity of active genes.
用微球菌核酸酶短暂消化鸡胚红细胞,会导致与β-珠蛋白序列相关的单体核小体以及β-珠蛋白基因侧翼的5'序列被优先切除并溶解。这两个区域在细胞核中对DNA酶I敏感。这种盐溶性核小体富含所有四种主要的HMG蛋白,但HMG1和2的结合较弱。这些核小体似乎已失去了活性基因的大部分DNA酶I敏感性。通过电泳分离的含HMG14和17的盐溶性核小体对DNA酶I不敏感,并且包含无活性基因序列以及活性序列。通过斑点印迹杂交分析,将HMG蛋白重组到大量核小体或染色质上未能揭示活性基因对HMG的依赖性敏感性,并且发现通过斑点印迹杂交分析的ASV前病毒序列的DNA酶I敏感性不依赖于HMG。这些结果表明,高级染色质结构可能是活性基因对核酸酶敏感性的原因。