Pavlovic J, Banz E, Parish R W
Institut für Immunologie und Virologie, Universität Zürich, Switzerland.
Nucleic Acids Res. 1989 Mar 25;17(6):2315-32. doi: 10.1093/nar/17.6.2315.
Micrococcal nuclease digestion of Dictyostelium discoideum nuclei from various developmental stages was used to investigate transcription-related changes in the chromatin structure of the coding region of four genes. Gene activity was determined by Northern blotting and nuclear run on experiments. During strong transcription of the developmentally regulated cysteine proteinase I gene, a smear superimposed on a nucleosomal ladder was observed, indicating perturbation of nucleosomal structure was occurring. However, two other developmentally regulated genes, discoidin I and pSC253, showed only slight nucleosome disruption during high levels of transcription. The chromatin structure of a fourth gene (pCZ22) was disrupted throughout development, even at those stages where transcription was greatly reduced. We suggest that although nucleosome structure can be transiently perturbed by the passage of the transcription complex in vivo, the degree of perturbation and the speed with which nucleosomes reassemble is also influenced by the DNA sequence.
利用微球菌核酸酶对不同发育阶段的盘基网柄菌细胞核进行消化,以研究四个基因编码区染色质结构中与转录相关的变化。通过Northern印迹法和细胞核连续转录实验确定基因活性。在发育调控的半胱氨酸蛋白酶I基因的强烈转录过程中,观察到在核小体梯状条带之上叠加有一条拖尾带,这表明核小体结构正在受到干扰。然而,另外两个发育调控基因,即盘状菌素I和pSC253,在高水平转录过程中仅表现出轻微的核小体破坏。第四个基因(pCZ22)的染色质结构在整个发育过程中都受到破坏,即使在转录大幅减少的阶段也是如此。我们认为,虽然转录复合物在体内通过时可使核小体结构暂时受到干扰,但干扰程度以及核小体重新组装的速度也受DNA序列的影响。