Bonner W M
Nucleic Acids Res. 1978 Jan;5(1):71-85. doi: 10.1093/nar/5.1.71.
Histone proximity in chromatin was studied with the cleavable crosslinking reagent, dithiobissuccinimidyl propionate. Crosslinks between H4 and H2a, H4 and H2b, H4 and H3, H2a and H2b, H2b and H3 were found. H1 is also crosslinked to the nucleosomal histones. In nuclei, unsheared chromatin, and H1 depleted chromatin, the four nucleosomal histones are crosslinked at similar relative rates both in 5 mM salt and 100 mM salt. After micrococcal nuclease treatment to generate nucleosomes, H2a and H2b are crosslinked faster than H4 and H3. C14-NEM titration of thiopropionate residues bound to each histone shows that H2a and H2b are more accessible to this reagent after nuclease treatment but that the increased binding was not sufficient by itself to explain the increase in crosslinking. Bolton Hunter reagent was used to further study the accessibility of the four nucleosomal histones in whole chromatin and nuclease digested chromatin. These studies showed that salt increases the accessibility of all four histones while nuclease treatment decreases H4 accessibility.
使用可裂解交联剂二硫代双琥珀酰亚胺丙酸酯研究染色质中的组蛋白接近度。发现H4与H2a、H4与H2b、H4与H3、H2a与H2b、H2b与H3之间存在交联。H1也与核小体组蛋白交联。在细胞核、未剪切的染色质和H1缺失的染色质中，在5 mM盐和100 mM盐中，四种核小体组蛋白以相似的相对速率交联。经微球菌核酸酶处理产生核小体后，H2a和H2b的交联速度比H4和H3快。对与每种组蛋白结合的硫代丙酸酯残基进行C14-NEM滴定表明，核酸酶处理后H2a和H2b对该试剂的可及性更高，但结合增加本身不足以解释交联增加。使用博尔顿·亨特试剂进一步研究全染色质和核酸酶消化染色质中四种核小体组蛋白的可及性。这些研究表明，盐增加了所有四种组蛋白的可及性，而核酸酶处理降低了H4的可及性。
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