Doudna J A, Szostak J W
Department of Molecular Biology, Massachusetts General Hospital, Boston 02114.
Mol Cell Biol. 1989 Dec;9(12):5480-3. doi: 10.1128/mcb.9.12.5480-5483.1989.
The self-splicing sunY intron from bacteriophage T4 has the smallest conserved core secondary structure of any of the active group I introns. Here we show that several nonconserved regions can be deleted from this intron without complete loss of catalytic activity. The 3' stems P9, P9.1, and P9.2 can be deleted while retaining 5' cleaving activity. Two base-paired stems (P7.1 and P7.2) that are peculiar to the group IA introns can also be deleted; however, the activities of the resulting derivatives depend greatly on the choice of replacement sequences and their lengths. The smallest active derivative is less than 180 nucleotides long. These experiments help to define the minimum structural requirements for catalysis.
来自噬菌体T4的自我剪接sunY内含子具有任何活性I组内含子中最小的保守核心二级结构。我们在此表明,该内含子的几个非保守区域可以被删除而不会完全丧失催化活性。3'茎P9、P9.1和P9.2可以被删除,同时保留5'切割活性。IA组内含子特有的两个碱基配对茎(P7.1和P7.2)也可以被删除;然而,所得衍生物的活性在很大程度上取决于替换序列的选择及其长度。最小的活性衍生物长度小于180个核苷酸。这些实验有助于确定催化的最低结构要求。