Chromatin condensation is confined to the loop and involves an all-or-none structural change

被引:13
作者
Balbi, C
Sanna, P
Barboro, P
Alberti, I
Barbesino, M
Patrone, E
机构
[1] Ist Nazl Ric Canc, I-16132 Genoa, Italy
[2] Inst Studi Chim Fis Macromol Sintet & Nat, I-16146 Genoa, Italy
关键词
D O I
10.1016/S0006-3495(99)77106-7
中图分类号
Q6 [生物物理学];
学科分类号
071011 ;
摘要
Using differential scanning calorimetry in combination with pulsed field gel electrophoresis; we relate here the changes in the thermal profile of rat liver nuclei induced by very mild digestion of chromatin by endogenous nuclease with the chain length distribution of the DNA fragments. The enthalpy of the endotherm at 106 degrees C, which reflects the denaturation of the heterochromatic domains, decreases dramatically after the induction of a very small number of double-strand breaks per chromosome; the thermal transition disappears when the loops have undergone on average one DNA chain scission event. Quantitative analysis of the experimental data shows that the loop behaves like a topologically isolated domain. Also discussed is the process of heterochromatin formation, which occurs according to an all-or-none mechanism. In the presence of spermine, a strong condensation agent, only the loops that have undergone one break are able Id refold, in confirmation of the extremely cooperative nature of the transition. Furthermore, our results suggest a-relationship between the states that give rise to the endotherms at 90 degrees C and 106 degrees C and the morphologies referred to as class II and dass III ina previous physicochemical study of the folding of chromatin fragments (Widom, 1986. J. Mel. Biol. 190:411-424) and support the view that the overall process of condensation follows a sequential (two-step) pathway.
引用
收藏
页码:2725 / 2735
页数:11
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