Ruffner D E, Dahm S C, Uhlenbeck O C
Department of Chemistry and Biochemistry, University of Colorado, Boulder 80309-0215.
Gene. 1989 Oct 15;82(1):31-41. doi: 10.1016/0378-1119(89)90027-9.
Nine different hammerhead RNA self-cleaving domains consistent with the consensus secondary structure proposed by Keese and Symons (1987) were prepared and tested for cleavage. Each hammerhead was constructed from two oligoribonucleotides in two different configurations. Although cleavage was observed in all nine cases, the rates of cleavage varied by more than a thousand fold. The presence of RNA secondary structure incompatible with hammerhead formation in the individual oligos may be responsible for the large rate differences. We have also examined the degree of participation of a proposed dimer hammerhead intermediate in one case and conclude that, while such a four-stranded structure can form, it is not the preferred reaction intermediate.
制备了与Keese和Symons（1987年）提出的共有二级结构一致的9种不同的锤头状RNA自我切割结构域，并对其切割活性进行了测试。每个锤头状结构由两个处于两种不同构型的寡核糖核苷酸构建而成。虽然在所有9种情况下均观察到了切割现象，但切割速率相差超过一千倍。各个寡核苷酸中存在与锤头状结构形成不相容的RNA二级结构可能是造成切割速率差异巨大的原因。我们还研究了在一个案例中所提出的二聚体锤头状中间体的参与程度，并得出结论：虽然这样的四链结构能够形成，但它并非首选的反应中间体。
