Barra J L, Rhounim L, Rossignol J L, Faugeron G
Institut Jacques Monod, UMR 7592, CNRS/Université Paris 7/Université Paris 6, 75251 Paris Cedex 05, France.
Mol Cell Biol. 2000 Jan;20(1):61-9. doi: 10.1128/MCB.20.1.61-69.2000.
A gene encoding a protein that shows sequence similarity with the histone H1 family only was cloned in Ascobolus immersus. The deduced peptide sequence presents the characteristic three-domain structure of metazoan linker histones, with a central globular region, an N-terminal tail, and a long positively charged C-terminal tail. By constructing an artificial duplication of this gene, named H1, it was possible to methylate and silence it by the MIP (methylation induced premeiotically) process. This resulted in the complete loss of the Ascobolus H1 histone. Mutant strains lacking H1 displayed normal methylation-associated gene silencing, underwent MIP, and showed the same methylation-associated chromatin modifications as did wild-type strains. However, they displayed an increased accessibility of micrococcal nuclease to chromatin, whether DNA was methylated or not, and exhibited a hypermethylation of the methylated genome compartment. These features are taken to imply that Ascobolus H1 histone is a ubiquitous component of chromatin which plays no role in methylation-associated gene silencing. Mutant strains lacking histone H1 reproduced normally through sexual crosses and displayed normal early vegetative growth. However, between 6 and 13 days after germination, they abruptly and consistently stopped growing, indicating that Ascobolus H1 histone is necessary for long life span. This constitutes the first observation of a physiologically important phenotype associated with the loss of H1.
仅与组蛋白H1家族表现出序列相似性的一种蛋白质编码基因在浸没状粪壳菌中被克隆出来。推导的肽序列呈现出后生动物连接组蛋白特有的三结构域结构,具有一个中央球状区域、一个N端尾巴和一个长的带正电荷的C端尾巴。通过构建这个名为H1的基因的人工重复序列,有可能通过减数分裂前甲基化诱导(MIP)过程使其甲基化并沉默。这导致浸没状粪壳菌H1组蛋白完全缺失。缺乏H1的突变菌株表现出正常的甲基化相关基因沉默,经历MIP过程,并显示出与野生型菌株相同的甲基化相关染色质修饰。然而,无论DNA是否甲基化,它们都表现出微球菌核酸酶对染色质的可及性增加,并且甲基化基因组区域出现超甲基化。这些特征意味着浸没状粪壳菌H1组蛋白是染色质的普遍组成部分,在甲基化相关基因沉默中不起作用。缺乏组蛋白H1的突变菌株通过有性杂交正常繁殖,并显示出正常的早期营养生长。然而,在萌发后6至13天之间,它们突然且持续停止生长,这表明浸没状粪壳菌H1组蛋白对长寿是必需的。这是首次观察到与H1缺失相关的具有生理重要性的表型。