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. Author manuscript; available in PMC: 2021 Jan 7.
Published in final edited form as: Cell Rep. 2020 Dec 22;33(12):108527. doi: 10.1016/j.celrep.2020.108527

Figure 5. RtcA Is Important for Operon Activation.

Figure 5.

(A–C) RNA from the indicated strains was subjected to northern blotting to detect 5′ halves of tRNATrp(CCA) (A), tRNALeu(UAG) (B), and tRNACys(GCA)

(C). In (A) and (B), specific fragments that become evident in Δrna strains are denoted by red lines in strains containing RNase I (lanes 5–8).

(D–F) After growing the indicated strains without or with MMC, RNA was subjected to northern blotting to detect 5′ halves of tRNAHis(GUG) (D), tRNALeu(UAG) (E) and tRNACys(GCA) (F).

(G) After growing the indicated FLAG3-rsr strains without or with MMC, lysates were subjected to immunoblotting to detect FLAG3-Rsr and RplE.

(H and I) Lysates of FLAG3-rsr strains carrying the indicated deletions were immunoblotted to detect FLAG3-Rsr. RplE, loading control. Right, quantitation (n = 3).

(J). Lysates of 14028s and SL1344 FLAG3-rsr strains carrying empty vector (lanes 3–5 and 9–11) or pHA-RtcA expressing RtcA fused to a hemagglutinin (HA) epitope tag (lane 6–8 and 12–14) were subjected to immunoblotting to detect FLAG3-Rsr, HA-RtcA, and RplE. Right, quantitation (n = 3). Data in (H), (I), and (J) are mean (n = 3) ± SEM. p values were calculated with two-tailed unpaired t tests. *p < 0.05, **p < 0.01, and ***p < 0.001; ns, not significant. See also Figures S4 and S5 and Table S2.