Marushige K, Marushige Y, Wong T K
Biochemistry. 1976 May 18;15(10):2047-53. doi: 10.1021/bi00655a004.
Displacement of histones from calf thymus chromatin has been studied in an attempt to postulate the mechanisms involved in the total removal of somatic-type histones during transformation of spermatid chromatin. When chromatin is saturated with protamine (protamine/DNA, 0.5), histone I becomes displaceable at 0.15-0.3 M NaCl, suggesting that direct replacement by highly basic sperm histone could be a mechanism for its removal. While histone I is the only histone which is extensively degraded upon incubation of chromatin and, therefore, proteolysis might provide an additional mechanism for the removal of this histone, acetylation of chromatin by acetic anhydride greatly increases suscpetibility of histones IIb1, IIb2, and III to the chromosomally associated protease. These histones are extensively degraded and displaced from the DNA upon incubation of the acetylated chromatin. Although histone IV is not appreciably degraded, the proteolytic removal of acetylated histone III from chromatin weakens the interaction of acetylated histone IV to the DNA, and this histone becomes dissociable at 0.3 M NaCl. A comparison of the extent of chemical acetylation of individual histones observed in this investigation with that of enzymatic acetylation which can be achieved in vivo suggests that acetylation and proteolysis could be a mechanism for the removal of histone IIb2 and III. The displacement of histones IIb1 and IV could be explained on the basis of decreased binding to DNA as a result of their acetylation together with the proteolytic removal of their respective partner histones, IIb2 and III.
为了推测精子细胞染色质转化过程中体细胞型组蛋白完全去除所涉及的机制,人们对小牛胸腺染色质中组蛋白的置换进行了研究。当染色质用鱼精蛋白饱和时(鱼精蛋白/DNA,0.5),组蛋白I在0.15 - 0.3 M NaCl浓度下变得可被置换,这表明由高度碱性的精子组蛋白直接替代可能是其去除的一种机制。虽然组蛋白I是染色质孵育时唯一大量降解的组蛋白,因此蛋白水解可能为该组蛋白的去除提供另一种机制,但用乙酸酐对染色质进行乙酰化会大大增加组蛋白IIb1、IIb2和III对染色体相关蛋白酶的敏感性。这些组蛋白在乙酰化染色质孵育时会大量降解并从DNA上被置换下来。虽然组蛋白IV没有明显降解,但从染色质中蛋白水解去除乙酰化的组蛋白III会削弱乙酰化组蛋白IV与DNA的相互作用,并且该组蛋白在0.3 M NaCl浓度下变得可解离。将本研究中观察到的单个组蛋白化学乙酰化程度与体内可实现的酶促乙酰化程度进行比较表明,乙酰化和蛋白水解可能是去除组蛋白IIb2和III的一种机制。组蛋白IIb1和IV的置换可以基于它们乙酰化导致与DNA结合减少以及它们各自的伴侣组蛋白IIb2和III被蛋白水解去除来解释。