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. Author manuscript; available in PMC: 2017 May 19.
Published in final edited form as: Cell. 2016 May 5;165(5):1171–1181. doi: 10.1016/j.cell.2016.04.018

Figure 3. The CS2a/TeSS domain is a P3-like domain in Tlc1.

Figure 3

(A) Schematic representation of the TeSS structure at the distal end of stem IVc of Tlc1 (left), the P3 domain of Nme1 RNase MRP RNA (middle) and the P3 domain of the Rpr1 RNase P RNA (right). Dark blue shading: identical nucleotides, light blue: purines and pyrimidines are conserved. Purple blue line on left of Tlc1: CS2a (Conserved Sequence element 2a; Gunisova et al., 2009). Dark green line on Nme1 RNA: nucleotides protected by Pop6/Pop7 binding (Perederina et al., 2007). For more details, see Figure S2A). (B) Telomere length analyses in strains harboring the indicated TLC1 alleles. TLC1: wild type; Δ: tlc1Δ; ΔL: tlc1-ΔL allele that lacks the TeSS/P3 domain (see Figure S4); P3NME1: tlc1::P3NME1, the TeSS/P3 domain in the Tlc1 RNA was replaced with the one from the Nme1 RNA; P3RPR1; tlc1::P3RPR1, the TeSS/P3 domain in Tlc1 was replaced with the one from the Rpr1 RNA. +: Strain carried a wt TLC1 gene on a URA3 plasmid. Black wedges indicate outgrowth of strains after loss of the plasmid borne TLC1 gene; last lane reflects growth for 110 generations. Schematic structures on top are color coded as the nucleotides in panel (A). (C) Growth assays of cells that contain the indicated NME1 alleles: nme1Δ, complete deletion of NME1; ΔP3: nme1-ΔP3 allele that lacks the P3 domain; nme1::P3TLC1 the NME1 P3 domain was replaced with the TLC1 TeSS/P3 domain. Bottom plate: all strains contained a wt NME1 gene on a plasmid with the URA3 marker. Top plate: growth of cells after loss of the URA3, NME1 containing plasmid. Schematic structures and color coding as in panel A.