RNA-modification by Base Exchange: Structure, Function and Application of tRNA-guanine Transglycosylases.

Abstract

tRNA-guanine transglycosylases (TGT) occur in all domains of life. They are unique among RNA-modifying enzymes as they exchange a guanine base in the primary RNA transcript by various 7-substituted 7-deazaguanines leading to the modified nucleosides queuosine and archaeosine. Archaeosine is found in the D-loop of archaeal tRNAs, queuosine in the anticodon of bacterial and eukaryotic tRNAs specific for Asp, Asn, His and Tyr. Structural and functional studies revealed a common base-exchange mechanism for all TGTs. Nonetheless, there are also significant differences between TGTs, which will be discussed here. It concerns the specificity for different 7-deazaguanine substrates as well as the recognition of substrate tRNAs. For queuosine TGT an anticodon stem-loop containing the UGU recognition motif is a minimal substrate sufficient for binding to the active site, however, full-length tRNA is bound with higher affinity due to multiple interactions with the dimeric enzyme. Archaeal TGT also binds tRNAs as homodimer, even though the interaction pattern is very different and results in a large change of tRNA conformation. Interestingly, a closely related enzyme, DpdA, exchanges guanine by 7-cyano-7-deazguanine (preQ₀) in double stranded DNA of several bacteria. Bacterial TGT is a target for structure-based drug design, as the virulence of Shigella depends on TGT activity, and mammalian TGT has been used for the treatment of murine experimental autoimmune encephalomyelitis, a model for chronic multiple sclerosis. Furthermore, TGT has become a valuable tool in nucleic acid chemistry, as it facilitates the incorporation of non-natural bases in tRNA molecules, e.g. for labelling or cross-linking purposes.