FAST FOLDING MUTANTS OF TETRAHYMENA RIBOZYME

进一步剖析四膜虫的折叠机制 核酶，我们正在与威廉姆森博士和 他的同事(目前在斯克里普斯研究所)，折叠 嗜热四膜虫I群的几个快速折叠突变体 内含子。这些突变特别令人感兴趣，因为它们是 都位于P4-P6领域，尽管它们深刻地影响着 核酶催化核心的折叠。威廉姆森博士 基团已经表征了折叠的温度和尿素依赖性 在这些突变体中，使用他们的寡核苷酸杂交方法， 它探测核酶中螺旋P3的形成。不管怎么说，这 HApproach无法探测可以遵循的快速过渡 Hby同步加速器足迹。为了测量 折叠其余400个碱基的Hof-Long RNA，Martha Rook，a 研究生Hin Williamsons实验室进行了初步的系列研究 这些突变体的时间分辨x射线足迹实验 与资源中心协作。

To further dissect the folding mechanism of the Tetrahymena ribozyme, we are exploring, in collaboration with Dr. Williamson and his co-workers (currently at Scripps Institute), the folding of several fast-folding mutants of the Tetrahymena thermophila group I intron. These mutations are of particular interest in that they are all located within the P4-P6 domain although they profoundly influence the folding of the catalytic core of the ribozyme. Dr. Williamsons group has characterized the temperature and urea dependence of folding Hof these mutants, using their oligonucleotide-hybridization method, Hwhich probes the formation of helix P3 in the ribozyme. owever, this Happroach is unable to probe the fast transitions that can be followed Hby synchrotron footprinting. In order to measure the rates of folding Hof the rest of the 400 bases-long RNA, Martha Rook, a graduate student Hin Williamsons lab conducted a preliminary series time-resolved x-ray footprinting experiments on these mutants in collaboration with the Resource Center.