Engineerability of a functional ribonucleoprotein complex

为了理解核糖核蛋白（RNP）结构和功能的分子基础，我们尝试了N抗终止复合物的相互作用和空间组织的工程。 RNA位点的空间方向的相对小的变化已被证明导致抗终止活性的大的变化。 我们发现，通过同时修饰RNA元件和N蛋白结构域的方向，可以部分恢复在改变抗终止复合物内核心HIV RRE位点的方向时失去的活性。 我们还表明，优化的N蛋白与NusA和RNA聚合酶的相互作用，导致抗终止活性的部分恢复。 结果表明，抗终止复合物内的核心相互作用的空间取向可以以合理的方式工程化，证明了核糖核蛋白复合物的工程化能力。

In order to understand the molecular basis for ribonucleoprotein (RNP) structure and function, we attempted the engineering of the interactions and the spatial organization of the N antitermination complex. Relatively small changes in the spatial orientation of the RNA site have been shown to result in large changes in antitermination activity. We found that it is possible to partially recover activity lost upon altering the orientation of the core HIV RRE site within the antitermination complex by simultaneously modifiyng the orientation of RNA elements and N protein domains. We also showed that optimization of the interaction of N protein with NusA and RNA polymerase lead to a partial recovery of antitermination activity. The results show that the spatial orientation of the core interaction within the antitermination complex can be engineered in a rational manner, demonstrating the engineerability of ribonucleoprotein complexes.