Fukuda Y
Molecular Biology Department, National Institute of Bioscience and Human Technology, AIST, MITI, Tsukuba, Ibaraki, Japan.
Plant Mol Biol. 2000 Sep;44(1):91-8. doi: 10.1023/a:1006416929665.
Two scaffold/matrix attachment regions (S/MARs), designated S/M I and S/M II, are located in the 5'-flanking region of the tobacco basic class I chitinase gene, CHN50. Structural analysis of these S/MARs showed that S/M II contained an intrinsically curved DNA sequence that is located between -1786 and -1722 relative to the initiation site of transcription. Electrophoretic mobility shift assays and southwestern blotting analysis were performed to identify the tobacco nuclear proteins that bind specifically to this curved DNA. These experiments revealed that nuclear proteins bound specifically to the curved DNA. Moreover, the nuclear proteins appeared to recognize the overall structure of the intrinsically curved DNA, as distinct from binding to the DNA with sequence specificity. Southwestern blotting analysis showed that proteins of 22, 24, 28 and 34 kDa bound specifically to the curved DNA. The possible functions of the binding proteins and their relationship to previously identified nuclear proteins, such as high-mobility-group proteins, are discussed.
两个支架/基质附着区域（S/MARs），分别命名为S/M I和S/M II，位于烟草I类基础几丁质酶基因CHN50的5'侧翼区域。对这些S/MARs的结构分析表明，S/M II包含一个内在弯曲的DNA序列，相对于转录起始位点，该序列位于-1786至-1722之间。进行了电泳迁移率变动分析和蛋白质免疫印迹分析，以鉴定与该弯曲DNA特异性结合的烟草核蛋白。这些实验表明，核蛋白与弯曲DNA特异性结合。此外，核蛋白似乎识别内在弯曲DNA的整体结构，这与序列特异性结合DNA不同。蛋白质免疫印迹分析表明，22 kDa、24 kDa、28 kDa和34 kDa的蛋白质与弯曲DNA特异性结合。文中讨论了结合蛋白的可能功能及其与先前鉴定的核蛋白（如高迁移率族蛋白）的关系。
