Williams S P, Langmore J P
Department of Biological Sciences, University of Michigan, Ann Arbor 48109-2099.
Biophys J. 1991 Mar;59(3):606-18. doi: 10.1016/S0006-3495(91)82276-7.
Analyses of low angle x-ray scattering from chromatin, isolated by identical procedures but from different species, indicate that fiber diameter and number of nucleosomes per unit length increase with the amount of nucleosome linker DNA. Experiments were conducted at physiological ionic strength to obtain parameters reflecting the structure most likely present in living cells. Guinier analyses were performed on scattering from solutions of soluble chromatin from Necturus maculosus erythrocytes (linker length 48 bp), chicken erythrocytes (linker length 64 bp), and Thyone briareus sperm (linker length 87 bp). The results were extrapolated to infinite dilution to eliminate interparticle contributions to the scattering. Cross-sectional radii of gyration were found to be 10.9 +/- 0.5, 12.1 +/- 0.4, and 15.9 +/- 0.5 nm for Necturus, chicken, and Thyone chromatin, respectively, which are consistent with fiber diameters of 30.8, 34.2, and 45.0 nm. Mass per unit lengths were found to be 6.9 +/- 0.5, 8.3 +/- 0.6, and 11.8 +/- 1.4 nucleosomes per 10 nm for Necturus, chicken, and Thyone chromatin, respectively. The geometrical consequences of the experimental mass per unit lengths and radii of gyration are consistent with a conserved interaction among nucleosomes. Cross-linking agents were found to have little effect on fiber external geometry, but significant effect on internal structure. The absolute values of fiber diameter and mass per unit length, and their dependencies upon linker length agree with the predictions of the double-helical crossed-linker model. A compilation of all published x-ray scattering data from the last decade indicates that the relationship between chromatin structure and linker length is consistent with data obtained by other investigators.
通过相同程序从不同物种中分离出的染色质进行的低角度X射线散射分析表明,纤维直径和每单位长度的核小体数量随核小体连接区DNA的量增加而增加。在生理离子强度下进行实验以获得反映活细胞中最可能存在的结构的参数。对黄斑美西螈红细胞(连接区长度48bp)、鸡红细胞(连接区长度64bp)和布里阿瑞俄斯海胆精子(连接区长度87bp)的可溶性染色质溶液的散射进行了吉尼埃分析。结果外推至无限稀释以消除颗粒间对散射的贡献。发现黄斑美西螈、鸡和海胆染色质的横截面回转半径分别为10.9±0.5、12.1±0.4和15.9±0.5nm,这与30.8、34.2和45.0nm的纤维直径一致。每10nm的单位长度质量分别为黄斑美西螈、鸡和海胆染色质的6.9±0.5、8.3±0.6和11.8±1.4个核小体。实验得到的单位长度质量和回转半径的几何结果与核小体之间保守的相互作用一致。发现交联剂对纤维外部几何形状影响很小,但对内部结构有显著影响。纤维直径和单位长度质量的绝对值及其对连接区长度的依赖性与双螺旋交联模型的预测一致。对过去十年所有已发表的X射线散射数据的汇编表明,染色质结构与连接区长度之间的关系与其他研究者获得的数据一致。