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.
Biophysics Program, Department of Radiology, Stanford University School of Medicine, 1201 Welch Road, Stanford, CA 94305-5484, USA.
Sumitaka
S
Hasegawa
Jianghong
J
Rao
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