Modulating the splicing activity of Tetrahymena ribozyme via RNA self-assembly.
The internal guiding sequence (IGS) is normally located at the 5' end of trans-splicing ribozymes that are derived from the Tetrahymena group I intron, and is required for the recognition of substrate RNAs and for trans-splicing reactions. Here, we separated the Tetrahymena group I intron at the L2 loop to produce two fragments: the IGS-containing substrate, and the IGS-lacking ribozyme. We show here that two fragments can complex not through the IGS interaction but under the guidance of appended interacting nucleotides, and perform trans-splicing. The splicing reactions took place both in vitro and in mammalian cells, and the spliced mRNA product from the self-assembled ribozyme complex can be translated into functional proteins in vivo. The splicing efficiency was dependent on the length of appending nucleotides.
Hasegawa
Sumitaka
S
Biophysics Program, Department of Radiology, Stanford University School of Medicine, 1201 Welch Road, Stanford, CA 94305-5484, USA.
Rao
Jianghong
J
D000818:Animals; D007438:Introns; D012326:RNA Splicing; D016337:RNA, Catalytic / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000737:chemistry / Q000378:metabolism*; D013768:Tetrahymena / Q000201:enzymology*
C116167:GIR1 ribozyme; D016337:RNA, Catalytic; D012333:RNA, Messenger