title: Promiscuous catalysis by the tetrahymena group I ribozyme.
abstract: Catalytic promiscuity, the ability of an enzyme to catalyze alternative reactions, has been suggested to have played an important role in the evolution of new catalytic activities in protein enzymes. Similarly, promiscuous activities may have been advantageous in an earlier RNA world. The Tetrahymena Group I ribozyme naturally catalyzes the site-specific guanosine attack on an anionic phosphate diester and has been shown to also catalyze aminoacyl transfer to water, albeit with a small rate acceleration (<10-fold). This inefficient catalysis could be due to the differences in charge and/or geometry requirements for the two reactions. Herein, we describe a new promiscuous activity of this ribozyme, the site-specific guanosine attack on a neutral phosphonate diester. This alternative substrate lacks the negative charge at the reaction center but, in contrast to the aminoacyl substrate, can undergo nucleophilic attack with the same geometry as the natural substrate. Our results show that the neutral phosphonate reaction is catalyzed about 1 x 106-fold, substantially better than the acyl transfer but far below the normal anionic substrate. We conclude that both charge and geometry are important factors for catalysis of the normal reaction and that promiscuous catalytic activities of ribozymes could have been created or enhanced by reorienting and swapping RNA domains.
authors.affiliation: Department of Biochemistry, B400 Beckman Center, Stanford University, Stanford, California 94305-5307, USA.
authors.forename: Marcello
authors.initials: M
authors.lastname: Forconi
authors.forename: Daniel
authors.initials: D
authors.lastname: Herschlag
mesh_terms: D000818:Animals; D002384:Catalysis; D006151:Guanosine / Q000378:metabolism; D007700:Kinetics; D063065:Organophosphonates / Q000378:metabolism; D016337:RNA, Catalytic / Q000235:genetics / Q000378:metabolism*; D013379:Substrate Specificity; D013768:Tetrahymena / Q000201:enzymology* / Q000235:genetics
chemical_list: D063065:Organophosphonates; D016337:RNA, Catalytic; D006151:Guanosine