de la Barre A E, Gerson V, Gout S, Creaven M, Allis C D, Dimitrov S
Laboratoire de Biologie Moléculaire et Cellulaire de la Différenciation, équipe Mécanismes d'Assemblage du Matériel Génétique, INSERM U 309, Institut Albert Bonniot, Domaine de la Merci, 38706 La Tronche, Cedex, France.
EMBO J. 2000 Feb 1;19(3):379-91. doi: 10.1093/emboj/19.3.379.
We have studied the role of core histone tails in the assembly of mitotic chromosomes using Xenopus egg extracts. Incubation of sperm nuclei in the extracts led to the formation of mitotic chromosomes, a process we found to be correlated with phosphorylation of the N-terminal tail of histone H3 at Ser10. When the extracts were supplemented with H1-depleted oligosomes, they were not able to assemble chromosomes. Selective elimination of oligosome histone tails by trypsin digestion resulted in a dramatic decrease in their ability to inhibit chromosome condensation. The chromosome assembly was also inhibited by each of the histone tails with differing efficiency. In addition, we found that nucleosomes were recruiting through the flexible histone tails some chromosome assembly factors, different from topoisomerase II and 13S condensin. These findings demonstrate that histone tails play an essential role in chromosome assembly. We also present evidence that the nucleosomes, through physical association, were able to deplete the extracts from the kinase phosphorylating histone H3 at Ser10, suggesting that this kinase could be important for chromosome condensation.
我们利用非洲爪蟾卵提取物研究了核心组蛋白尾巴在有丝分裂染色体组装中的作用。将精子细胞核在提取物中孵育会导致有丝分裂染色体的形成,我们发现这一过程与组蛋白H3 N端尾巴在丝氨酸10处的磷酸化相关。当提取物中添加了去除H1的寡核小体时,它们无法组装染色体。用胰蛋白酶消化选择性去除寡核小体的组蛋白尾巴,导致其抑制染色体凝聚的能力显著下降。不同组蛋白尾巴对染色体组装的抑制效率也各不相同。此外,我们发现核小体通过灵活的组蛋白尾巴招募了一些不同于拓扑异构酶II和13S凝聚素的染色体组装因子。这些发现表明组蛋白尾巴在染色体组装中起着至关重要的作用。我们还提供了证据表明,核小体通过物理结合能够从在丝氨酸10处磷酸化组蛋白H3的激酶中耗尽提取物,这表明这种激酶可能对染色体凝聚很重要。