Parseghian M H, Newcomb R L, Winokur S T, Hamkalo B A
Research and Development, Techniclone Corporation, Tustin, CA 92780, USA.
Chromosome Res. 2000;8(5):405-24. doi: 10.1023/a:1009262819961.
Chromatin immunoprecipitation was employed to determine whether or not the previously reported depletion of histone H1 on actively transcribed sequences was selective with respect to H1 subtypes. DNA of immunofractionated chromatin was analyzed by slot-blots for repetitive sequences and PCR for single and low-copy sequences. Based on the analysis of a diverse set of sequences, we report distinct differences in subtype distributions. Actively transcribed chromatin, as well as chromatin poised for transcription, is characterized by a relative depletion of somatic H1 subtypes 2 and 4 (H1s-2 and H1s-4),whereas facultative and constitutive heterochromatin contain all four somatic subtypes. These results support a model in which subtypes are selectively depleted upon gene expression. In turn, the data also support the possibility that the somatic subtypes have different functional roles based on their selective depletion from different classes of DNA sequences.
采用染色质免疫沉淀法来确定先前报道的活跃转录序列上组蛋白H1的缺失是否对H1亚型具有选择性。通过狭缝印迹分析免疫分离染色质的DNA以检测重复序列,并通过PCR分析单拷贝和低拷贝序列。基于对多种序列的分析,我们报道了亚型分布的明显差异。活跃转录的染色质以及准备转录的染色质的特征是体细胞H1亚型2和4(H1s-2和H1s-4)相对缺失,而兼性和组成型异染色质包含所有四种体细胞亚型。这些结果支持了一个模型,即基因表达时亚型会被选择性耗尽。反过来,这些数据也支持了这样一种可能性,即基于体细胞亚型从不同类别的DNA序列中被选择性耗尽,它们具有不同的功能作用。