Horvath M P, Schultz S C
Department of Chemistry and Biochemistry, University of Colorado, Boulder, CO 80309-0215, USA.
J Mol Biol. 2001 Jul 6;310(2):367-77. doi: 10.1006/jmbi.2001.4766.
The Oxytricha nova telomere end binding protein (OnTEBP) recognizes, binds and protects the single-stranded 3'-terminal DNA extension found at the ends of macronuclear chromosomes. The structure of this complex shows that the single strand GGGGTTTTGGGG DNA binds in a deep cleft between the two protein subunits of OnTEBP, adopting a non-helical and irregular conformation. In extending the resolution limit of this structure to 1.86 A, we were surprised to find a G-quartet linked dimer of the GGGGTTTTGGGG DNA also packing within the crystal lattice and interacting with the telomere end binding protein. The G-quartet DNA exhibits the same structure and topology as previously observed in solution by NMR with diagonally crossing d(TTTT) loops at either end of the four-stranded helix. Additionally, the crystal structure reveals clearly visible Na(+), and specific patterns of bound water molecules in the four non-equivalent grooves. Although the G-quartet:protein contact surfaces are modest and might simply represent crystal packing interactions, it is interesting to speculate that the two types of telomeric DNA-protein interactions observed here might both be important in telomere biology.
新大核草履虫端粒末端结合蛋白(OnTEBP)能够识别、结合并保护在大核染色体末端发现的单链3'-末端DNA延伸。该复合物的结构表明,单链GGGGTTTTGGGG DNA结合在OnTEBP两个蛋白质亚基之间的一个深裂隙中,呈非螺旋且不规则的构象。将该结构的分辨率极限扩展到1.86 Å时,我们惊讶地发现,GGGGTTTTGGGG DNA的一个G-四联体连接二聚体也堆积在晶格中,并与端粒末端结合蛋白相互作用。G-四联体DNA呈现出与之前在溶液中通过核磁共振观察到的相同结构和拓扑结构,在四链螺旋的两端有对角交叉的d(TTTT)环。此外,晶体结构清晰地显示出可见的Na(+),以及在四个不等价沟中结合水分子的特定模式。尽管G-四联体与蛋白质的接触表面较小,可能仅仅代表晶体堆积相互作用,但有趣的是,可以推测这里观察到的两种端粒DNA-蛋白质相互作用可能在端粒生物学中都很重要。