Tse C, Fletcher T M, Hansen J C
Department of Biochemistry, University of Texas Health Science Center, 7703 Floyd Curl Drive, San Antonio, TX 78284-7760, USA.
Proc Natl Acad Sci U S A. 1998 Oct 13;95(21):12169-73. doi: 10.1073/pnas.95.21.12169.
Defined model systems consisting of physiologically spaced arrays of H3/H4 tetramer.5S rDNA complexes have been assembled in vitro from pure components. Analytical hydrodynamic and electrophoretic studies have revealed that the structural features of H3/H4 tetramer arrays closely resemble those of naked DNA. The reptation in agarose gels of H3/H4 tetramer arrays is essentially indistinguishable from naked DNA, the gel-free mobility of H3/H4 tetramer arrays relative to naked DNA is reduced by only 6% compared with 20% for nucleosomal arrays, and H3/H4 tetramer arrays are incapable of folding under ionic conditions where nucleosomal arrays are extensively folded. We further show that the cognate binding sites for transcription factor TFIIIA are significantly more accessible when the rDNA is complexed with H3/H4 tetramers than with histone octamers. These results suggest that the processes of DNA replication and transcription have evolved to exploit the unique structural properties of H3/H4 tetramer arrays.
由生理间距的H3/H4四聚体.5S rDNA复合物阵列组成的特定模型系统已在体外由纯组分组装而成。分析性流体动力学和电泳研究表明,H3/H4四聚体阵列的结构特征与裸露DNA的结构特征极为相似。H3/H4四聚体阵列在琼脂糖凝胶中的蠕动与裸露DNA基本无法区分,H3/H4四聚体阵列相对于裸露DNA的无凝胶迁移率仅降低了6%,而核小体阵列则降低了20%,并且H3/H4四聚体阵列在核小体阵列广泛折叠的离子条件下无法折叠。我们进一步表明,当rDNA与H3/H4四聚体复合时,转录因子TFIIIA的同源结合位点比与组蛋白八聚体复合时更容易接近。这些结果表明,DNA复制和转录过程已经进化到利用H3/H4四聚体阵列的独特结构特性。