Identification and Sequence Analysis of Metazoan tRNA 3′-End Processing Enzymes tRNase Zs

Abstract

tRNase Z is the endonuclease responsible for removing the 3'-trailer sequences from precursor tRNAs, a prerequisite for the addition of the CCA sequence. It occurs in the short (tRNase ZS) and long (tRNase ZL) forms. Here we report the identification and sequence analysis of candidate tRNase Zs from 81 metazoan species. We found that the vast majority of deuterostomes, lophotrochozoans and lower metazoans have one tRNase ZS and one tRNase ZL genes, whereas ecdysozoans possess only a single tRNase ZL gene. Sequence analysis revealed that in metazoans, a single nuclear tRNase ZL gene is likely to encode both the nuclear and mitochondrial forms of tRNA 3′-end processing enzyme through mechanisms that include alternative translation initiation from two in-frame start codons and alternative splicing. Sequence conservation analysis revealed a variant PxKxRN motif, PxPxRG, which is located in the N-terminal region of tRNase ZSs. We also identified a previously unappreciated motif, AxDx, present in the C-terminal region of both tRNase ZSs and tRNase ZLs. The AxDx motif consisting mainly of a very short loop is potentially close enough to form hydrogen bonds with the loop containing the PxKxRN or PxPxRG motif. Through complementation analysis, we demonstrated the likely functional importance of the AxDx motif. In conclusion, our analysis supports the notion that in metazoans a single tRNase ZL has evolved to participate in both nuclear and mitochondrial tRNA 3′-end processing, whereas tRNase ZS may have evolved new functions. Our analysis also unveils new evolutionarily conserved motifs in tRNase Zs, including the C-terminal AxDx motif, which may have functional significance.