The structural basis of RNA-catalyzed RNA polymerization

Early life presumably required polymerase ribozymes capable of replicating RNA. Known polymerase ribozymes best approximating such replicases use as their catalytic engine an RNA-ligase ribozyme originally selected from random RNA sequences. Here, we report 3.15 Å crystal structures of this ligase trapped in catalytically viable pre-ligation states, with the 3′-hydroxyl nucleophile positioned for in-line attack on the 5′-triphosphate. Guided by metal and solvent-mediated interactions, the 5′-triphosphate hooks into the major groove of the adjoining RNA duplex in an unanticipated conformation. Two phosphates and the nucleophile jointly coordinate an active-site metal ion. Atomic mutagenesis experiments demonstrate that active-site nucleobase and hydroxyl groups also participate directly in catalysis, collectively playing a role that in proteinaceous polymerases is performed by a second metal ion. Thus artificial ribozymes can employ complex catalytic strategies that differ dramatically from those of analogous biological enzymes. The RNA world hypothesis proposes that primordial organisms lacked both DNA genes and protein enzymes, and instead used RNA as the sole biopolymer of information-storage an