Maier Verena K, Chioda Mariacristina, Rhodes Daniela, Becker Peter B
Molekularbiologie, Adolf-Butenandt Institut, Ludwig Maximilian Universität, Munich, Germany.
EMBO J. 2008 Mar 19;27(6):817-26. doi: 10.1038/sj.emboj.7601902. Epub 2007 Oct 25.
Nucleosome-remodelling factors containing the ATPase ISWI, such as ACF, render DNA in chromatin accessible by promoting the sliding of histone octamers. Although the ATP-dependent repositioning of mononucleosomes is readily observable in vitro, it is unclear to which extent nucleosomes can be moved in physiological chromatin, where neighbouring nucleosomes, linker histones and the folding of the nucleosomal array restrict mobility. We assembled arrays consisting of 12 nucleosomes or 12 chromatosomes (nucleosomes plus linker histone) from defined components and subjected them to remodelling by ACF or the ATPase CHD1. Both factors increased the access to DNA in nucleosome arrays. ACF, but not CHD1, catalysed profound movements of nucleosomes throughout the array, suggesting different remodelling mechanisms. Linker histones inhibited remodelling by CHD1. Surprisingly, ACF catalysed significant repositioning of entire chromatosomes in chromatin containing saturating levels of linker histone H1. H1 inhibited the ATP-dependent generation of DNA accessibility by only about 50%. This first demonstration of catalysed chromatosome movements suggests that the bulk of interphase euchromatin may be rendered dynamic by dedicated nucleosome-remodelling factors.
含有ATP酶ISWI的核小体重塑因子,如ACF,通过促进组蛋白八聚体的滑动使染色质中的DNA易于接近。尽管在体外很容易观察到单核小体的ATP依赖重新定位,但尚不清楚在生理染色质中核小体能在多大程度上移动,在生理染色质中,相邻的核小体、连接组蛋白和核小体阵列的折叠限制了其移动性。我们从确定的组分组装了由12个核小体或12个染色质体(核小体加连接组蛋白)组成的阵列,并使其接受ACF或ATP酶CHD1的重塑。这两种因子都增加了核小体阵列中DNA的可及性。ACF而非CHD1催化了整个阵列中核小体的深度移动,表明存在不同的重塑机制。连接组蛋白抑制CHD1的重塑作用。令人惊讶的是,在含有饱和水平连接组蛋白H1的染色质中,ACF催化了整个染色质体的显著重新定位。H1仅使ATP依赖的DNA可及性的产生抑制约50%。对催化染色质体移动的首次证明表明,大部分间期常染色质可能通过专门的核小体重塑因子而变得具有动态性。