Wu H N, Lee J Y, Ping Y H, Wang C N
Institute of Molecular Biology, Academia Sinica, Taipei, Taiwan, ROC.
FEBS Lett. 1998 Nov 20;439(3):312-6. doi: 10.1016/s0014-5793(98)01395-7.
Oligonucleotides were inserted into the Lp3 region of a hepatitis delta virus (HDV) cis-cleaving ribozyme and the effect of Lp3 enlargement on ribozyme folding was examined by assaying the activity of each mutant. The location of insertion and the sequence of the inserted oligonucleotide had distinct effects on ribozyme activity, and the HDV ribozyme was capable of adopting the active structure for cis-cleavage when a pentanucleotide was inserted into the 3' portion of Lp3. Furthermore, whether the insertion mutant cis-cleaved or not, the structure of Lp3 was altered by the inserted oligonucleotide. These findings disclose that the 5' rather than the 3' portion of Lp3 is critical for ribozyme folding and catalysis.
将寡核苷酸插入丁型肝炎病毒（HDV）顺式切割核酶的Lp3区域，并通过检测每个突变体的活性来研究Lp3扩大对核酶折叠的影响。插入位置和插入寡核苷酸的序列对核酶活性有不同影响，当将五核苷酸插入Lp3的3'部分时，HDV核酶能够采用顺式切割的活性结构。此外，无论插入突变体是否进行顺式切割，插入的寡核苷酸都会改变Lp3的结构。这些发现表明，Lp3的5'部分而非3'部分对核酶折叠和催化至关重要。
