Pemov A Iu, Hamlin D L, Bavykin S G
Department of Biochemistry, University of Virginia School of Medicine, Charlottesville 22908, USA.
Mol Biol (Mosk). 1995 Jul-Aug;29(4):832-40.
We investigated histone-DNA interactions in chromatin of amplified about 1000 fold dihydrofolate reductase (DHFR) domain in methotrexate resistant Chinese hamster ovary cell line CHOC 400. We explored chromatin structure of DHFR-gene and extended region lying 3'-downstream of the DHFR-gene, which includes two zones of DNA replication initiation, Ori-beta and Ori-gamma, and matrix attachment region (MAR). Using the method of hybridization with "protein images" we found that the yield of both core histones and histone H1 cross-linked with DNA in 5'-region of the DHFR-gene, in Ori-gamma and regions flanked from either side MAR is significantly decreased in comparison with 3'-transcribed region of the DHFR-gene, Ori-beta and MAR. It was shown previously that decreased yield of histones cross-linked with DNA is typical for 5'-region of actively transcribed genes. So, histone-DNA interactions in chromatin of the 5'-region of the house-keeping and moderately transcribed DHFR-gene, Ori-gamma and regions which flank MAR resemble those in 5'-regions of actively transcribed genes. Significant differences in amount of histones cross-linked with DNA in various parts of the DHFR-domain testify to high level of heterogeneity of chromatin structure in the domain.
我们研究了甲氨蝶呤抗性中国仓鼠卵巢细胞系CHOC 400中扩增约1000倍的二氢叶酸还原酶(DHFR)结构域染色质中的组蛋白-DNA相互作用。我们探索了DHFR基因的染色质结构以及位于DHFR基因3'下游的延伸区域,该区域包括两个DNA复制起始区,即Ori-β和Ori-γ,以及基质附着区域(MAR)。使用与“蛋白质图像”杂交的方法,我们发现与DHFR基因5'区域、Ori-γ以及MAR两侧区域中的DNA交联的核心组蛋白和组蛋白H1的产量,与DHFR基因的3'转录区域、Ori-β和MAR相比显著降低。先前已表明,与DNA交联的组蛋白产量降低是活跃转录基因5'区域的典型特征。因此,管家基因和中度转录的DHFR基因5'区域、Ori-γ以及MAR两侧区域染色质中的组蛋白-DNA相互作用类似于活跃转录基因5'区域中的相互作用。DHFR结构域不同部位与DNA交联的组蛋白数量存在显著差异,这证明了该结构域染色质结构的高度异质性。