Article
- The EMBO Journal (2001) 20, 5453 - 5460
- doi:10.1093/emboj/20.19.5453
Design and development of a catalytic ribonucleoprotein
Shota Atsumi1, Yoshiya Ikawa1,2, Hideaki Shiraishi1,2 and Tan Inoue1,2
- Graduate School of Science, Kyoto University, Kyoto 606-8502, Japan
- Graduate School of Biostudies, Kyoto University, Kyoto 606-8502, Japan
Correspondence to:
Tan Inoue, E-mail: tan@kuchem.kyoto-u.ac.jp
Received 9 September 2000; Accepted 15 August 2001; Revised 3 August 2001
Abstract
Ribonucleoproteins (RNPs) consisting of derivatives of a ribozyme and an RNA-binding protein were designed and constructed based upon high-resolution structures of the corresponding prototype molecules, the Tetrahymena group I self-splicing intron RNA and two proteins (bacteriophage 
N and HIV Rev proteins) containing RNA-binding motifs. The splicing reaction proceeds efficiently only when the designed RNA associates with the designed protein either in vivo or in vitro. In vivo mutagenic protein selection was effective for improving the capability of the protein. Kinetic analyses indicate that the protein promotes RNA folding to establish an active conformation. The fact that the conversion of a ribozyme to an RNP can be accomplished by simple molecular design supports the RNA world hypothesis and suggests that a natural active RNP might have evolved readily from a ribozyme.
Keywords:
- group I intron,
- in vivo selection,
- ribonucleoprotein,
- ribozyme,
- RNA–protein interaction
