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. 2021 Nov 18;17(11):e1009899. doi: 10.1371/journal.pgen.1009899

Fig 1. TPR orthologs in yeast contribute to tRNA export.

Fig 1

(A) Binding of Nup60 at tRNA genes. ChIP-qPCR of Nup60-TAP was performed with strains MC177 (wt), MC273 (Δmlp1), MC276 (Δmlp1,2), and MC172 (no TAP) after M phase arrest by CDC20 depletion, when Nup60 was known to have increased association with tRNA genes in the wt strain. Data (mean ± SD, n = 3) were analyzed by Student’s t-test in paired comparison with the same tRNA gene in the wt strain; *P < 0.05. (B) tRNA gene-NPC association. Strains MC199 [tS(CGA)C::256lacop], MC200 [tT(UGU)G1::256lacop], MRG7425 [tS(CGA)C::256lacop Δmlp1,2], and MRG7426 [tT(UGU)G1::256lacop Δmlp1,2] were examined after M-phase arrest. Colocalization was defined as GFP and RFP foci were separated by no more than the width of the GFP spot. Scale bar = 10 μm. N denotes the number of cells analysed. Pairwise χ2-tests, *P < 0.05. (C) Nascent tRNA levels. RT-qPCR was done with strains MC177 (wt) and MC276 (Δmlp1,2) after arrest in M phase to measure the levels of three representative pre-tRNAs and nascent ACT1 mRNA. Values were normalized to mature ACT1 mRNA. (D) Western blot to detect the expression of Gcn4-TAP in asynchronously grown cultures of strains MC341 (wt), MC343 (Δlos1), MC349 (Δnup2), MC350 (Δnup60), MC344 (Δmlp1) and MC345 (Δmlp1,2). Coomassie blue staining was used to show even amount of protein loaded in gels.