Miyashita T, Yamamoto H, Takemoto Y, Nozaki M, Morita T, Matsushiro A
Department of Microbial Genetics, Osaka University, Japan.
Gene. 1993 Mar 30;125(2):151-8. doi: 10.1016/0378-1119(93)90322-t.
DNase I hypersensitive (DH) sites in a 12-kb genomic fragment carrying the mouse EndoA gene were examined to obtain information on the changes in chromatin structure associated with activation of this gene encoding extra-endodermal cytoskeletal protein A (EndoA) during early mouse embryogenesis. Seven DH sites were found in this locus in parietal yolk-sac-like cells, PYS-2, which produce EndoA constitutively. In differentiated mouse teratocarcinoma F9 cells that produce EndoA inductively, this locus has three DH sites. In both cell lines, these sites were mapped to the upstream region of the promoter, the promoter and the 3' enhancer region. The DNA of PYS-2 cells has one more DH site within the first exon and three additional DH sites within the first intron. These DH sites are not present in DNA from BALB/c 3T3 cells and undifferentiated F9 cells that do not produce EndoA. Thus, the formation of these differentiation-dependent DH sites is required for the differentiation-specific expression of the mouse EndoA. In addition, another strong DH site, which may be associated with the B2 element expression, was detected in the third intron of the gene in undifferentiated F9 cells.
对携带小鼠EndoA基因的12 kb基因组片段中的DNase I超敏(DH)位点进行了检测,以获取有关在小鼠早期胚胎发育过程中与编码内胚层外细胞骨架蛋白A(EndoA)的该基因激活相关的染色质结构变化的信息。在组成性产生EndoA的卵黄囊样细胞PYS-2中的该基因座中发现了7个DH位点。在诱导产生EndoA的分化小鼠畸胎瘤F9细胞中,该基因座有3个DH位点。在这两种细胞系中,这些位点都定位于启动子的上游区域、启动子和3'增强子区域。PYS-2细胞的DNA在第一个外显子内还有一个DH位点,在第一个内含子内还有另外3个DH位点。这些DH位点不存在于不产生EndoA的BALB/c 3T3细胞和未分化的F9细胞的DNA中。因此,这些依赖分化的DH位点的形成是小鼠EndoA分化特异性表达所必需的。此外,在未分化的F9细胞的基因的第三个内含子中检测到另一个可能与B2元件表达相关的强DH位点。