The participation of poly(ADP-ribosyl)ated histone H1 in oligonucleosomal condensation.
The participation of poly(ADP-ribosyl)ated histone H1 in oligonucleosomal condensation.
复制标题
聚（ADP-核糖基）化组蛋白 H1 参与寡核小体缩合。
DOI：
10.1111/j.1432-1033.1982.tb06953.x
复制
发表时间：
1982
期刊：
影响因子：
--
通讯作者：
Smulson,M
中科院分区：
文献类型：
--
作者：
Wong,M;Malik,N;Smulson,M
染色质相关酶聚（ADP-Rib）聚合酶引起修饰的寡核小体的NAD依赖性交联，如通过电泳和沉降分析所证明的[Butt，T. R.和Smulson，M.（1980）Biochemistry，19，5235-5242]。据推测，组蛋白H1的聚（ADP-核糖基）化和随后通过交联形成H1二聚体可能是这种现象的重要组成部分。为了研究这一过程，需要一种将组蛋白H1与染色质复合的方法，以促进该组蛋白的准确聚（ADP-核糖基）化的恢复。之前，我们已经确定两个组蛋白H1分子通过长度为15或16个单元的聚（ADP‐Rib）链交联。在当前的研究中，我们利用用H1复溶的寡核小体的能力来合成聚（ADP‐Rib）-H1复合物，以监测复溶的准确性。看来，一个特定的距离和相邻的H1分子沿沿着的多核小体纤维的酶促合成这种修改的组蛋白complex.We建立了一个受控的胰蛋白酶消化oligonucleosomes删除H1组蛋白与其他核蛋白与染色质的干扰最小。此外，通过该程序从染色质中部分去除了聚（ADP-Rib）聚合酶。随后，利用梯度盐透析的方法将聚合酶和组蛋白H1重组到耗尽的寡核小体中，通过上述方法完成了H1（和聚合酶）与寡核小体内特异性结合位点的再结合。在耗尽的寡核小体中未观察到聚（ADP-Rib）-H1-二聚体合成，但发现这种能力在重建的染色质中部分恢复。类似地，NAD促进核小体交联的能力在重构样品中恢复。这些结果为进一步研究组蛋白的聚腺苷二磷酸核糖基化如何改变染色质结构提供了基础。
The chromatin‐associated enzyme poly(ADP‐Rib) polymerase causes an NAD‐dependent crosslinking of modified oligonucleosomes, as demonstrated by electrophoretic and sedimentation analysis [Butt, T. R. and Smulson, M. (1980)Biochemistry, 19, 5235–5242]. It was speculated that poly(ADP‐ribosyl)ation of histone H1 and subsequent formation through crosslinking to an H1 dimer may be an important component of this phenomenon. To study this process, a method of complexing histone H1 to chromatin was required that promoted the restoration of accurate poly(ADP‐ribosyl)ation of this histone. Previously we have established that two histone H1 molecules are crosslinked by a chain of poly(ADP‐Rib) 15 or 16 units in length. In the current study, we made use of the ability of oligonucleosomes, reconstituted with H1, to carry out the synthesis of the poly(ADP‐Rib)–H1 complex in order to monitor the accuracy of reconstitution. It appears that a specific distance and juxtaposition of adjacent H1 molecules along the polynucleosome fiber is required for the enzymatic synthesis of this modified histone complex.We established that a controlled trypsin digestion of oligonucleosomes removed H1 histone with minimal perturbation of other nuclear proteins associated with chromatin. In addition, poly(ADP‐Rib) polymerase was partially removed from chromatin by this procedure. Subsequently, methods utilizing gradient salt dialysis have been employed to reconstitute both the polymerase and histone H1 to the depleted oligonucleosomes.The reassociation of H1 (and polymerase) to specific binding sites within oligonucleosomes was accomplished by the above procedures. Poly(ADP‐Rib)–H1‐dimer synthesis was not observed in depleted oligonucleosomes, but this capacity was found to be partially restored in the reconstituted chromatin. Similiarly, the ability of NAD to promote crosslinking of nucleosomes was restored in the reconstituted samples. These results provide a basis for further studies on how the poly(ADP‐ribosyl)ation of histones alters the structure of chromatin.