Görisch Sabine M, Wachsmuth Malte, Tóth Katalin Fejes, Lichter Peter, Rippe Karsten
Division of Molecular Genetics, Deutsches Krebsforschungszentrum, Im Neuenheimer Feld 280, 69120 Heidelberg, Germany.
J Cell Sci. 2005 Dec 15;118(Pt 24):5825-34. doi: 10.1242/jcs.02689. Epub 2005 Nov 29.
In eukaryotes, the interaction of DNA with proteins and supramolecular complexes involved in gene expression is controlled by the dynamic organization of chromatin inasmuch as it defines the DNA accessibility. Here, the nuclear distribution of microinjected fluorescein-labeled dextrans of 42 kDa to 2.5 MDa molecular mass was used to characterize the chromatin accessibility in dependence on histone acetylation. Measurements of the fluorescein-dextran sizes were combined with an image correlation spectroscopy analysis, and three different interphase chromatin condensation states with apparent pore sizes of 16-20 nm, 36-56 nm and 60-100 nm were identified. A reversible change of the chromatin conformation to a uniform 60-100 nm pore size distribution was observed upon increased histone acetylation. This result identifies histone acetylation as a central factor in the dynamic regulation of chromatin accessibility during interphase. In mitotic chromosomes, the chromatin exclusion limit was 10-20 nm and independent of the histone acetylation state.
在真核生物中,DNA与参与基因表达的蛋白质及超分子复合物之间的相互作用受染色质动态组织的控制,因为它决定了DNA的可及性。在此,利用显微注射的分子量为42 kDa至2.5 MDa的荧光素标记葡聚糖在细胞核中的分布来表征依赖于组蛋白乙酰化的染色质可及性。荧光素 - 葡聚糖大小的测量与图像相关光谱分析相结合,鉴定出三种不同的间期染色质凝聚状态,其表观孔径分别为16 - 20 nm、36 - 56 nm和60 - 100 nm。随着组蛋白乙酰化增加,观察到染色质构象可逆地转变为均匀的60 - 100 nm孔径分布。该结果表明组蛋白乙酰化是间期染色质可及性动态调节的核心因素。在有丝分裂染色体中,染色质排斥极限为10 - 20 nm,且与组蛋白乙酰化状态无关。