The Early Apoptotic DNA Fragmentation Targets a Small Number of Specific Open Chromatin Regions
The Early Apoptotic DNA Fragmentation Targets a Small Number of Specific Open Chromatin Regions
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DOI：
10.1371/journal.pone.0005010
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发表时间：
2009-04-06
期刊：
影响因子：
3.7
通讯作者：
Bernardi, Giorgio
中科院分区：
文献类型：
--
作者：
Di Filippo, Miriam;Bernardi, Giorgio
我们在这里报告，早期凋亡的DNA片段，通过使用一种全新的方法，是在少数特定的开放染色质区域的间期核攻击的结果。这被证明如下：（i）切除鸡肝并在37 ℃下在无菌管中保持1至3小时;（ii）这诱导了细胞凋亡。（可能是由于缺氧），如DNA制备物产生的电泳核小体梯所示;（iii）低分子量DNA片段（类似于200 bp）进行克隆、测序，并显示主要来源于基因和周围100 kb区域;（iv）产生几百个切口，通常涉及相同的染色体位点;（v）在可比的DNA降解水平下，微球菌核酸酶（MNase）也显示出对基因和周围区域的一般偏好，但MNase切割位于与以下位点完全不同的位点，并且比通过凋亡切割的那些更不特异。总之，这里提出的方法，这是最温和和最少的侵入性的方法，确定了一个首选的可访问性景观间期染色质。
We report here that early apoptotic DNA fragmentation, as obtained by using an entirely new approach, is the result of an attack at a small number of specific open chromatin regions of interphase nuclei. This was demonstrated as follows: (i) chicken liver was excised and kept in sterile tubes for 1 to 3 hours at 37 degrees C; (ii) this induced apoptosis (possibly because of oxygen deprivation), as shown by the electrophoretic nucleosomal ladder produced by DNA preparations; (iii) low molecular-weight DNA fragments (similar to 200 bp) were cloned, sequenced, and shown to derive predominantly from genes and surrounding 100 kb regions; (iv) a few hundred cuts were produced, very often involving the same chromosomal sites; (v) at comparable DNA degradation levels, micrococcal nuclease (MNase) also showed a general preference for genes and surrounding regions, but MNase cuts were located at sites that were quite distinct from, and less specific than, those cut by apoptosis. In conclusion, the approach presented here, which is the mildest and least intrusive approach, identifies a preferred accessibility landscape in interphase chromatin.