Engelke D R, Gottesfeld J M
Department of Biological Chemistry, University of Michigan, Ann Arbor 48109.
Nucleic Acids Res. 1990 Oct 25;18(20):6031-7. doi: 10.1093/nar/18.20.6031.
The chromatin structure of the Xenopus oocyte-specific 5S rRNA genes was examined at high resolution in immature oocyte and somatic cell chromosomes by DNase I footprinting. On oocyte chromatin, where the genes are active, the cleavage preferences over the entire gene region showed a periodic pattern of sensitivity and were dramatically different from the patterns obtained with deproteinized DNA or somatic cell chromatin. Further, the normal binding site for TFIIIA over the internal promoter region was preferentially sensitive to cleavage, indicating that TFIIIA was not bound in the manner predicted by in vitro experiments. In somatic cell chromatin, the oocyte-type 5S genes displayed a cleavage pattern largely similar to deproteinized DNA suggesting the absence of positioned nucleosomes on these inactive genes, although the presence of uncharacterized repressor complexes could not be ruled out. These data are discussed in terms of potential forms of the chromatin structure and alternative mechanisms of oocyte-type gene activation.
通过DNase I足迹法在高分辨率下检测了非洲爪蟾卵母细胞特异性5S rRNA基因在未成熟卵母细胞和体细胞染色体中的染色质结构。在基因活跃的卵母细胞染色质上,整个基因区域的切割偏好呈现出周期性的敏感模式,与用脱蛋白DNA或体细胞染色质获得的模式显著不同。此外,TFIIIA在内含启动子区域的正常结合位点对切割优先敏感,这表明TFIIIA并非以体外实验预测的方式结合。在体细胞染色质中,卵母细胞型5S基因呈现出与脱蛋白DNA大致相似的切割模式,这表明这些无活性基因上不存在定位核小体,尽管不能排除存在未鉴定的阻遏复合物。根据染色质结构的潜在形式和卵母细胞型基因激活的替代机制对这些数据进行了讨论。