Department of Chemistry and Biochemistry, Florida International University, 11200 SW 8th Street, Miami, Florida 33199, USA.
Biochemistry. 2010 Mar 2;49(8):1590-5. doi: 10.1021/bi901881c.
The mammalian high-mobility group protein AT hook 2 (HMGA2) is a DNA binding protein that specifically recognizes the minor groove of AT-rich DNA sequences. Disruption of its expression pattern is directly linked to oncogenesis and obesity. In this paper, we constructed a new plasmid pBendAT to study HMGA2-induced DNA bending. pBendAT carries a 230 bp DNA segment containing five pairs of restriction enzyme sites, which can be used to produce a set of DNA fragments of identical length to study protein-induced DNA bending. The DNA fragments of identical length can also be generated using PCR amplification. Since pBendAT does not contain more than three consecutive AT base pairs, it is suitable for the assessment of DNA bending induced by proteins recognizing AT-rich DNA sequences. Indeed, using pBendAT, we demonstrated that HMGA2 is a DNA bending protein and bends all three tested DNA binding sequences of HMGA2, SELEX1, SELEX2, and PRDII. The DNA bending angles were estimated to be 34.2 degrees , 33.5 degrees , and 35.4 degrees , respectively.
哺乳动物高迁移率族蛋白 A2（HMGA2）是一种 DNA 结合蛋白，它能特异性识别富含 AT 的 DNA 序列的小沟。其表达模式的破坏与肿瘤发生和肥胖直接相关。在本文中，我们构建了一个新的质粒 pBendAT 来研究 HMGA2 诱导的 DNA 弯曲。pBendAT 携带一个包含 5 对限制酶位点的 230 bp DNA 片段，可用于产生一组长度相同的 DNA 片段，以研究蛋白诱导的 DNA 弯曲。使用 PCR 扩增也可以生成长度相同的 DNA 片段。由于 pBendAT 不包含超过三个连续的 AT 碱基对，因此它适用于评估识别富含 AT 的 DNA 序列的蛋白诱导的 DNA 弯曲。事实上，使用 pBendAT，我们证明了 HMGA2 是一种 DNA 弯曲蛋白，并弯曲了所有三个测试的 HMGA2 的 DNA 结合序列，SELEX1、SELEX2 和 PRDII。DNA 弯曲角度分别估计为 34.2 度、33.5 度和 35.4 度。
