Department of Biotechnology, Graduate School of Engineering, Nagoya University, Furo-cho, Chikusa-ku, Nagoya, 464-8603, Japan.
Cytotechnology. 2006 Nov;52(3):159-70. doi: 10.1007/s10616-006-9017-4. Epub 2006 Sep 26.
Chicken lysozyme is highly expressed in the oviduct. The 5' regulatory region of this gene contains a negative element that represses transcription. To assess the molecular basis underlying the regulation of lysozyme gene expression, we investigated the binding protein to this region. Sequence motif analysis suggested the existence of putative YY1 binding sites in this regulatory region. Electrophoretic mobility shift assay showed the specific binding of YY1 to the negative element. In addition, chromatin immunoprecipitation assay indicated that YY1 specifically bound to the negative element in oviduct cells but not in erythrocytes. It was suggested by electrophoretic mobility shift assay and chromatin immunoprecipitation assay that YY1 also bound to the negative regulatory region in the promoter of the ovalbumin gene which also shows oviduct-specific expression. Western blot analysis showed that YY1 was expressed in relatively high levels in the oviduct and nucleus fractionation experiments showed that YY1 was localized both in chromosome and nuclear matrix fractions. These results suggest that there are some specific roles in the negative regulatory regions of these genes in relation to the multifunctional transcription factor YY1.
鸡溶菌酶在输卵管中高度表达。该基因的 5'调控区含有一个负调控元件,抑制转录。为了评估溶菌酶基因表达调控的分子基础,我们研究了该区域的结合蛋白。序列模体分析表明,在这个调控区域存在潜在的 YY1 结合位点。电泳迁移率变动分析显示 YY1 与负调控元件特异性结合。此外,染色质免疫沉淀分析表明,YY1 特异性结合于输卵管细胞中的负调控元件,而不是在红细胞中。电泳迁移率变动分析和染色质免疫沉淀分析表明,YY1 也与同样具有输卵管特异性表达的卵清蛋白基因启动子的负调控区结合。Western blot 分析表明,YY1 在输卵管中表达水平相对较高,核分数化实验表明,YY1 定位于染色体和核基质分数中。这些结果表明,在这些基因的负调控区域中,存在一些与多功能转录因子 YY1 相关的特定作用。