Nickel B E, Davie J R
Department of Biochemistry, Faculty of Medicine, University of Manitoba, Winnipeg, Canada.
Biochim Biophys Acta. 1989 Jan 23;1007(1):23-9. doi: 10.1016/0167-4781(89)90125-5.
Micrococcal nuclease was used to probe the nucleosomal organization of the rainbow trout germ-line-specific protamine multi-gene family in testis and erythrocytes. In erythrocyte chromatin, the repressed protamine genes show a distinct nucleosomal repeat pattern. However, in early-stage testis chromatin, where the protamine genes are expressed, they lack a distinct nucleosomal repeat pattern, indicating that the disrupted chromatin structure is related to their transcriptional activity. Micrococcal nuclease-digested testis and erythrocyte chromatin was separated into soluble and insoluble fractions. Transcriptionally active/competent genes of testis that had been labeled by nuclear nick-translation were enriched in the low-salt eluted, micrococcal nuclease-sensitive chromatin fraction. This fraction was not enriched in protamine DNA sequences. In testis, but not erythrocytes, protamine DNA sequences were slightly enriched in chromatin that fractionated with insoluble nuclear material, suggesting that transcriptionally active protamine gene chromatin has an insoluble character. Since the different protamine genes may not be simultaneously expressed, our results show the distribution of both transcriptionally active and inactive protamine genes. However, our observations indicate that the active germ-line-specific protamine gene chromatin shares several, but not all, of the features associated with other active tissue-specific genes.
使用微球菌核酸酶来探测虹鳟鱼生殖系特异性鱼精蛋白多基因家族在睾丸和红细胞中的核小体组织。在红细胞染色质中,受抑制的鱼精蛋白基因呈现出独特的核小体重复模式。然而,在鱼精蛋白基因表达的早期睾丸染色质中,它们缺乏明显的核小体重复模式,这表明染色质结构的破坏与其转录活性有关。微球菌核酸酶消化的睾丸和红细胞染色质被分离成可溶和不可溶部分。通过核切口平移标记的睾丸转录活性/有转录能力的基因在低盐洗脱的、对微球菌核酸酶敏感的染色质部分中富集。该部分在鱼精蛋白DNA序列中未富集。在睾丸而非红细胞中,鱼精蛋白DNA序列在与不溶性核物质一起分级分离的染色质中略有富集,这表明转录活性的鱼精蛋白基因染色质具有不溶性特征。由于不同的鱼精蛋白基因可能不会同时表达,我们的结果显示了转录活性和非活性鱼精蛋白基因的分布。然而,我们的观察表明,活跃的生殖系特异性鱼精蛋白基因染色质具有一些但并非所有与其他活跃的组织特异性基因相关的特征。