Selections for constituting new RNA-protein interactions in catalytic RNP.
In vitro and in vivo selection techniques are developed to constitute new RNA-peptide interactions. The selection strategy is designed by employing a catalytic RNP consisting of a derivative of the Tetrahymena ribozyme and an artificial RNA-binding protein. An arginine-rich RNA-binding motif and its target RNA motif in the RNP are substituted with randomized sequences and used for the selection experiments. Previously unknown binding motifs are obtained and the newly established interactions have been indispensable for assembling a catalytically active RNP. The method employed in this study is useful for making customized self-splicing intron RNAs whose activity is regulated by protein cofactors.
Atsumi
Shota
S
Graduate School of Science, Kyoto University, Kyoto 606-8502, Japan.
Ikawa
Yoshiya
Y
Shiraishi
Hideaki
H
Inoue
Tan
T
D001665:Binding Sites / Q000235:genetics; D024202:Electrophoretic Mobility Shift Assay; D015694:Gene Products, rev / Q000235:genetics / Q000378:metabolism; D010957:Plasmids / Q000235:genetics; D011485:Protein Binding; D012313:RNA / Q000378:metabolism*; D012326:RNA Splicing; D016337:RNA, Catalytic / Q000235:genetics* / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000235:genetics* / Q000378:metabolism; D012261:Ribonucleoproteins / Q000235:genetics* / Q000378:metabolism
D015694:Gene Products, rev; D016337:RNA, Catalytic; D016601:RNA-Binding Proteins; D012261:Ribonucleoproteins; D012313:RNA