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离子强度和温度升高诱导的染色质结构重排。

Rearrangement of chromatin structure induced by increasing ionic strength and temperature.

作者信息

Spadafora C, Oudet P, Chambon P

出版信息

Eur J Biochem. 1979 Oct;100(1):225-35. doi: 10.1111/j.1432-1033.1979.tb02053.x.

Abstract

Native rat liver chromatin fragments exposed to 600 mM NaCl at 37 degrees C for 45 min exhibit substantial modification of their original (approximately 200 base pairs) repeating subunit structure: a new repeat of 140 base pairs, superimposed on a high background, is observed after micrococcal nuclease digestion. The same material appears, in the electron microscope, as clusters of tightly packed beads connected by stretches of 'free' DNA. These modifications are not observed when the native chromatin is incubated at 37 degrees C at NaCl concentrations up to 400 mM. When native rat liver chromatin depleted of histone H1 by tRNA extraction is exposed to ionic strengths up to 600 mM NaCl at 4 degrees C, almost no modifications of the original native repeating structure are observed. However, when the incubation is carried out at 37 degrees C in 150, 300 or 400 mM NaCl, rearrangements of the native structure occur as indicated by micrococcal nuclease digestion and electron microscopic studies. Incubation of H1-depleted chromatin at 600 mM NaCl for 45 min at 37 degrees C induces, as for the native chromatin, a complete rearrangement characterized by the appearance of a 140-base-pair repeat superimposed on a high background upon digestion by micrococcal nuclease. It is suggested that these rearrangements are mediated by hydrophobic interactions between the histone cores and are prevented at ionic strengths lower than 500 mM by the presence of histone H1.

摘要

将原代大鼠肝脏染色质片段在37℃下于600 mM NaCl中孵育45分钟后,其原始(约200个碱基对)的重复亚基结构会发生显著改变:经微球菌核酸酶消化后,可观察到一个140个碱基对的新重复序列叠加在高背景之上。在电子显微镜下,同样的物质呈现为由“游离”DNA片段连接的紧密堆积的珠子簇。当原代染色质在37℃下于NaCl浓度高达400 mM的环境中孵育时,未观察到这些改变。当通过tRNA提取去除组蛋白H1的原代大鼠肝脏染色质在4℃下暴露于高达600 mM NaCl的离子强度时,几乎未观察到原始天然重复结构的改变。然而,当在37℃下于150、300或400 mM NaCl中孵育时,如微球菌核酸酶消化和电子显微镜研究所示,天然结构会发生重排。在37℃下将去除H1的染色质在600 mM NaCl中孵育45分钟,与原代染色质一样,会诱导完全重排,其特征是经微球菌核酸酶消化后出现叠加在高背景上的140个碱基对的重复序列。有人提出,这些重排是由组蛋白核心之间的疏水相互作用介导的,并且在离子强度低于500 mM时,由于组蛋白H1的存在而受到抑制。

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