Figure 2.

Rnt1 is involved in rancRNA_18 biogenesis. (A) secondary structure prediction of the 5 ’end of the TRM10 mRNA with the rancRNA_18 sequence indicated in red and a downstream AGNN tetraloop in orange. (B) growth analysis of a tetracycline (tet)-inducible rnt1 depletion strain (tet_off) in the presence (orange) or absence (green) of doxycycline compared to wildtype (wt, black and blue). n = 6, mean ± SD (C) northern blot analysis on total RNA isolated from wt, trm10Δ, xrn1Δ, rrp6Δ and Rnt1::tet cells either in the absence (-tet) or presence (+tet) of tetracycline. the locations of rancRNA_18 and the putative 49 nucleotides long precursor are indicated. the ethidium-bromide stained tRNAs serve as a loading control. (D) growth of the tetraloop mutant strain (orange) compared to wildtype (black) and trm10Δ (blue) strains was monitored either in normal (left) or high salt media. as a complementation control the trm10Δ strain transformed with a plasmid containing the wt TRM10 locus (green). n = 3, mean ± SD