5′-End sequencing in Saccharomyces cerevisiae offers new insights into 5′-ends of tRNAHis and snoRNAs

Abstract

tRNAHis guanylyltransferase (Thg1) specifies eukaryotic tRNAHis identity by catalysing a 3′–5′ non-Watson–Crick (WC) addition of guanosine to the 5′-end of tRNAHis. Thg1 family enzymes in Archaea and Bacteria, called Thg1-like proteins (TLPs), catalyse a similar but distinct 3′–5′ addition in an exclusively WC-dependent manner. Here, a genetic system in Saccharomyces cerevisiae was employed to further assess the biochemical differences between Thg1 and TLPs. Utilizing a novel 5′-end sequencing pipeline, we find that a Bacillus thuringiensis TLP sustains the growth of a thg1Δ strain by maintaining a WC-dependent addition of U−1 across from A73. Additionally, we observe 5′-end heterogeneity in S. cerevisiae small nucleolar RNAs (snoRNAs), an observation that may inform methods of annotation and mechanisms of snoRNA processing.