Varshavsky A J, Nedospasov S A, Schmatchenko V V, Bakayev V V, Chumackov P M, Georgiev G P
Nucleic Acids Res. 1977 Oct;4(10):3303-25. doi: 10.1093/nar/4.10.3303.
We report two new findings bearing on the "supranucleo-somal" level of the structure of the Simian Virus 40 minichromosome. I) Isolated SV40 minichromosome which contains all five histones including HI/I/ exists in solution under approximately physiological ionic conditions as a compact roughly spherical particle approximately 300 A in diameter which is capable of fitting within the virus capsid. In spite of such a compact conformation of the minichromosome individual nucleosomes can be readily visualized within the particle. Compact state of SV40 minichromosome depends on both the presence of histone HI and maintenance of approximately physiological ionic strength of solution (micron approximately 0.15). Removal of HI results in a conversion of the compact minichromosomes into an extended (circular beaded) structure. 2) The compact form of the SV40 minichromosome in contract to its circular beaded form is virtually completely resistant to staphylococcal nuclease, strongly suggesting that in particular nuclease-sensitive parts of the internucleosomal DNA regions are not exposed on the outside of the compact SV40 minichromosome. On the other hand, DNase I which is known to attack both inter-and intranucleosomal DNA in the chronatin /2,3/ readily digests the compact form of the SV40 minichromosome. Possible models of the compact minichromosome and implications for higher order structures of the cellular chromatin are discussed.
我们报告了两项关于猴病毒40(SV40)微型染色体结构“核小体以上”水平的新发现。1)分离出的包含所有五种组蛋白(包括H1)的SV40微型染色体,在大约生理离子条件下的溶液中以直径约300埃的紧密大致球形颗粒形式存在,该颗粒能够装入病毒衣壳内。尽管微型染色体具有如此紧密的构象,但在颗粒内仍可轻易观察到单个核小体。SV40微型染色体的紧密状态取决于组蛋白H1的存在以及溶液中大约生理离子强度的维持(离子强度约为0.15)。去除H1会导致紧密的微型染色体转变为伸展的(圆形串珠状)结构。2)与圆形串珠状形式相比,SV40微型染色体的紧密形式几乎完全抵抗葡萄球菌核酸酶,这强烈表明核小体间DNA区域中特别对核酸酶敏感的部分在紧密的SV40微型染色体外部并未暴露。另一方面,已知能攻击染色质中核小体间和核小体内DNA的DNase I能轻易消化SV40微型染色体的紧密形式。文中讨论了紧密微型染色体的可能模型及其对细胞染色质更高阶结构的影响。